Composition comprising casein and method for producing the same

文档序号：441517 发布日期：2021-12-28 浏览：13次中文

阅读说明：本技术 包含酪蛋白的组合物及其生产方法 (Composition comprising casein and method for producing the same ) 是由 R.潘迪亚 P.甘地 S.纪 D.博尚 L.洪于 2015-08-21 设计创作，主要内容包括：本申请涉及包含酪蛋白的组合物及其生产方法。本文公开了包括酪蛋白的方法和组合物,以及制备这些组合物的方法。(The present application relates to compositions comprising casein and methods for producing the same. Disclosed herein are methods and compositions comprising casein, and methods of making these compositions.)

1. A substitute dairy composition, wherein:

(a) the substitute dairy composition comprises one or more identified recombinant milk proteins;

(b) the substitute dairy composition is substantially free of milk proteins other than the one or more identified recombinant milk proteins; and

i. a recombinant beta-lactoglobulin protein which is capable of producing,

recombinant beta-lactoglobulin protein and recombinant alpha-lactalbumin protein,

recombinant beta-lactoglobulin protein and recombinant kappa-casein protein, or

Recombinant beta-lactoglobulin proteins and recombinant beta-casein proteins.

2. The substitute dairy composition of claim 1, wherein the one or more identified recombinant milk proteins are substantially free of milk impurities.

3. The substitute dairy composition of claim 1, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

4. The substitute dairy composition of claim 1, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

5. The substitute dairy composition of claim 1, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

6. The substitute dairy composition of claim 1, wherein at least one of the one or more identified recombinant dairy proteins is produced by a fungal cell or a bacterial cell.

7. The substitute dairy composition of claim 6, wherein said fungal cells are Aspergillus or Trichoderma.

8. The substitute dairy composition of claim 1, further comprising one or more of: calcium, potassium, sodium, citrate, thiamine (vitamin B1), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), vitamin B6 (pyridoxine), vitamin B12 (cobalamin), vitamin C, folate, vitamin a, vitamin D, vitamin E and vitamin K.

9. The substitute dairy composition of claim 1, further comprising one or more sweeteners selected from the group consisting of brazzein, curculin, mabinlin, miraculin, monellin, betadine, and thaumatin.

10. The substitute dairy composition of claim 1, further comprising one or more lipids derived from a plant selected from the group consisting of: soybean, sunflower, coconut, peanut, cottonseed, olive, palm, rapeseed, safflower, sesame seed, soybean, almond, beech nut, brazil nut, cashew, hazelnut, macadamia nut, mongolian nut, pecan, pine nut, pistachio nut, walnut, avocado, corn, canola, safflower, linseed, palm kernel, palm fruit, babassu, shea fruit, mango, cocoa, wheat and rice.

11. The substitute dairy composition of claim 1, further comprising one or more monoglycerides, diglycerides, triglycerides and/or phospholipids comprising short chain fatty acids selected from butyric acid, caproic acid, caprylic acid and capric acid.

12. The substitute dairy composition of claim 11, wherein at least one of the one or more monoglycerides, diglycerides, triglycerides, and/or phospholipids is a plant-based trans-esterified monoglyceride, diglyceride, triglyceride, and/or phospholipid.

13. The substitute dairy composition of claim 11, wherein the one or more monoglycerides, diglycerides, triglycerides, and/or phospholipids comprise fatty acyl chains in a percentage similar to the percentage of fatty acyl chains found in milk or dairy products of mammalian origin.

14. The substitute dairy composition of claim 1, wherein the substitute dairy composition has improved storage stability compared to a corresponding dairy composition.

15. The substitute dairy composition of claim 1, wherein the substitute dairy composition comprises a plurality of micelles comprising recombinant β -lactoglobulin protein, recombinant α -lactalbumin protein, recombinant κ -casein protein, and/or recombinant β -casein protein.

16. The substitute dairy composition of claim 1, wherein the substitute dairy composition comprises a polymer matrix gel comprising recombinant β -lactoglobulin protein and/or recombinant α -lactalbumin protein.

17. The substitute dairy composition of claim 1, wherein said substitute dairy composition is selected from the group consisting of ice cream, yogurt, cheese, frozen custard, creme fraiche, curd, cottage cheese, cream cheese, buttermilk, and butter.

18. The substitute dairy composition of claim 1, wherein the substitute dairy composition is a powder composition.

19. A substitute dairy composition, wherein:

(a) the replacement dairy composition comprises a mammal-produced milk or a mammal-produced dairy fraction, and a fraction comprising one or more identified recombinant milk proteins;

(b) the substitute dairy composition is substantially free of any recombinant milk protein other than the one or more identified recombinant milk proteins; and

i. a recombinant beta-lactoglobulin protein which is capable of producing,

recombinant beta-lactoglobulin protein and recombinant alpha-lactalbumin protein,

Recombinant beta-lactoglobulin protein and recombinant kappa-casein protein, or

Recombinant beta-lactoglobulin proteins and recombinant beta-casein proteins.

20. The substitute dairy composition of claim 19, wherein the one or more identified recombinant milk proteins are substantially free of milk impurities.

21. The substitute dairy composition of claim 19, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 80% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

22. The substitute dairy composition of claim 19, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

23. The substitute dairy composition of claim 19, wherein at least one of the one or more identified recombinant milk proteins comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of a bovine milk protein, a sheep milk protein, a horse milk protein, or a goat milk protein.

24. The substitute dairy composition of claim 19, wherein at least one of the one or more identified recombinant dairy proteins is produced by a fungal cell or a bacterial cell.

25. The substitute dairy composition of claim 24, wherein said fungal cells are aspergillus or trichoderma.

26. The substitute dairy composition of claim 19, further comprising one or more of: calcium, potassium, sodium, citrate, thiamine (vitamin B1), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), vitamin B6 (pyridoxine), vitamin B12 (cobalamin), vitamin C, folate, vitamin a, vitamin D, vitamin E and vitamin K.

27. The substitute dairy composition of claim 19, wherein the substitute dairy composition has improved storage stability compared to a corresponding dairy composition.

28. The substitute dairy composition of claim 19, wherein the substitute dairy composition comprises a plurality of micelles comprising recombinant β -lactoglobulin protein, recombinant α -lactalbumin protein, recombinant κ -casein protein, and/or recombinant β -casein protein.

29. The substitute dairy composition of claim 19, wherein the substitute dairy composition comprises a polymer matrix gel comprising recombinant β -lactoglobulin protein and/or recombinant α -lactalbumin protein.

30. The substitute dairy composition of claim 19, wherein said substitute dairy composition is an ice cream substitute, a yogurt substitute, a cheese substitute, a frozen custard substitute, a french creme substitute, a curd substitute, a cottage cheese substitute, a cream cheese substitute, a buttermilk substitute, or a butter substitute.

Technical Field

The present invention relates to a dairy substitute, a method of manufacturing thereof and a composition comprising milk fat free of animal ingredients and proteins for food applications such as milk, butter, cheese, yoghurt and cream.

Background

The global dairy market is estimated to be $ 5000 billion with an average annual growth rate of 4%. Cow milk accounts for a large portion of the market, while plant-based alternatives account for $ 10 billion in the united states, and an estimated $ 7 billion for lactose-intolerant milk. Bovine milk is known to have four specific caseins, α -s 1-casein, α -s 2-casein, β -casein and κ -casein. Milk produced by mammals or mammalians is a very complex fluid that includes thousands of components (e.g., if all triglycerides are identified). Mammals or mammals produce Milk that includes water, various lipids, sugars, various Proteins, and various inorganic salts and compounds (see, e.g., bland and Thompson (Eds), Milk Proteins from Expression to Food, Academic Press, 2014). Although milk produced by mammals, such as cow's milk, is considered by many to be an ideal nutritional source, various milk substitutes, such as plant or nut based milks, for example soy, almond or coconut milk, for milk produced by mammals, such as cow's milk, have been sought for reasons related to allergenicity of milk produced by mammals or mammals, lactose intolerance of certain components, personal preference, and perceived reduced environmental benefits of the dairy industry.

For example, environmental effects caused by dairy emissions can result in significant levels of nitrate, which is likely to contaminate groundwater. Groundwater forms the main water supply source for many towns and farms where surface water supply is limited. In the united states, half of the population relies entirely or partially on ground water, while europe has similar data (see, for example, the victoria environment and primary industry government websites at www.depi.vic.gov.au/aggregation-and-food/day/gaming-effluent/day-effluent-protecting-group water). The presence of food-borne pathogens in milk is due to direct contact with contaminated sources in dairy farm environments and to the udder excretions of infected animals. Outbreaks of human disease have traced to consumption of unpasteurized milk and also to pasteurized milk. The major contaminants commonly encountered in milk and dairy products include pesticide residues, heavy metals and aflatoxin M1(Awasthi et al,Indian J. Public Health 56:95-99, 2012)。

existing dairy substitutes, such as soy, almond or coconut milk, do not meet standards in terms of flavor and function; furthermore, most industrial and cultural significance of dairy milk derives from its usefulness in derived products such as cheese, yoghurt, cream or butter. Non-dairy plant based milk, while addressing environmental and health concerns (and while providing adequate flavor to a small portion of the population), it is almost universally impossible to form such derivative products when subjected to the same processes used for dairy milk.

Therefore, what is needed is a dairy substitute or composition having desirable flavor and performance characteristics, for example, a composition that replicates dairy flavor, minimizes food-borne pathogens, and has a lower environmental impact in production, while retaining derivatives or downstream applications for dairy milk and while providing nutritional characteristics similar to mammals or milk produced by mammals.

Summary of The Invention

The present invention is based on the following findings: only a subset of the components of the mammalian-produced milk may be used to generate compositions having similar flavor, similar appearance, similar nutritional value, similar aroma, and similar mouthfeel of the mammalian-produced milk.

Provided herein are compositions comprising: from about 0.3g/L to about 1.1g/L of kappa-casein protein; about 1.25g/L to about 4.9g/L of beta-casein protein; (ii) one or more lipids at a final total concentration of about 0 wt% to about 45 wt%; one or more flavor compounds at a final total concentration of about 0.01% to about 6% by weight; one or more sweeteners in a final total concentration of about 0.1% to about 6% by weight; and ash at a final total concentration of about 0.15 wt% to about 1.5 wt%, wherein the composition does not include animal-derived components.

Also provided are compositions comprising: from about 0.3g/L to about 1.1g/L of kappa-casein protein; about 1.25g/L to about 4.9g/L of beta-casein protein; (ii) one or more lipids at a final total concentration of about 0 wt% to about 45 wt%; one or more flavor compounds at a final total concentration of about 0.01% to about 6% by weight; one or more sweeteners in a final total concentration of about 0.1% to about 6% by weight; and ash at a final total concentration of about 0.15 wt% to about 1.5 wt%, wherein the composition: excluding at least one component found in milk produced by a mammal; comprising at least one component not present in mammalian-produced milk; and/or comprises at least one component at a higher or lower concentration than the concentration of at least one component in milk produced by the mammal. In some embodiments of these compositions, the composition comprises a higher concentration of at least one component compared to the concentration of one or more components in milk produced by the mammal, the component selected from the group consisting of: calcium, phosphate, B complex vitamin, vitamin A, vitamin D, vitamin E and vitamin K. In some embodiments of these compositions, the composition does not include at least one component found in milk produced by a mammal selected from the group consisting of: lactose, bacteria, mycobacteria, allergens, viruses, prions, yeast, growth hormones, leukocytes, antibiotics, heavy metals, immunoglobulins, lactoferrin, lactoperoxidase and lipase. In some embodiments of these compositions, wherein the composition comprises at least one component not present in mammalian-produced milk, the component is selected from the group consisting of artificial sweeteners, plant-derived lipids, β -casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern, and κ -casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern.

Also provided are compositions comprising: from about 0.3g/L to about 1.1g/L of a kappa-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern; from about 1.25g/L to about 4.9g/L of β -casein protein that is not glycosylated or has a non-mammalian glycosylation pattern; (ii) one or more lipids at a final total concentration of about 0 wt% to about 45 wt%; one or more flavor compounds at a final total concentration of about 0.01% to about 6% by weight; one or more sweeteners in a final total concentration of about 0.1% to about 6% by weight; and ash in a final total concentration of about 0.15 wt% to about 1.5 wt%.

Also provided are compositions comprising micelles comprising kappa-casein protein and beta-casein protein, wherein the micelles have a diameter of about 50nm to about 350nm, and the kappa-casein protein and beta-casein protein are non-glycosylated or have a non-mammalian glycosylation pattern. In some embodiments of these methods, the composition comprises micelles at a final concentration of about 2.0 wt.% to about 6 wt.%. In some embodiments of these compositions, the ratio of beta-casein protein to kappa-casein protein in the micelle is about 3.5:1 to about 5.5:1 (e.g., about 4:1 to about 5: 1). In some embodiments of these methods, the composition further comprises: (ii) one or more lipids at a final total concentration of about 0 wt% to about 45 wt%; one or more flavor compounds at a final total concentration of about 0.01% to about 6% by weight; one or more sweeteners in a final total concentration of about 0.1% to about 6% by weight; and ash in a final total concentration of about 0.15 wt% to about 1.5 wt%.

In some embodiments of any of the compositions described herein, the composition comprises from about 0.27% to about 0.75% by weight kappa-casein protein and from about 1.23% to about 3.27% by weight beta-casein protein. In some embodiments of any of the compositions described herein, the final total concentration of the one or more lipids is from about 0 wt.% to about 4.5 wt.%.

In some embodiments of any of the compositions described herein, the flavor compounds comprise at least one flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone. In some embodiments of any of the compositions described herein, the one or more sweeteners are sugar. In some embodiments of any of the compositions described herein, the saccharide is selected from the group consisting of: glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. In some embodiments of any of the compositions described herein, the one or more sweeteners are artificial sweeteners. In some embodiments of any of the compositions described herein, the artificial sweetener is selected from the group consisting of: stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol (lactitol), mabinlin, maltitol, mannitol, miraculin, monatin, monellin, ostwald (osladin), betadine, sorbitol, thaumatin, xylitol, potassium acetamidosulfonate, advatame, alitame, aspartame-acesulfame, cyclamate, dulcin, neohesperidin dihydrochalcone, neotame, and P-4000.

In some embodiments of any of the compositions described herein, the ash comprises one or more of: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some embodiments of any of the compositions described herein, the ash comprises CaCl₂、KH₂PO₄And sodium citrate (e.g., one, two, or three). In some embodiments of any of the compositions described herein, the CaCl₂Has a final concentration of about 0.05g/L to about 0.2 g/L; KH (Perkin Elmer)₂PO₄Has a final concentration of about 0.2g/L to about 0.4 g/L; and the sodium citrate has a final concentration of about 0.1g/L to about 0.3 g/L.

In some embodiments of any of the compositions described herein, the kappa-casein protein is a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth kappa-casein protein. In some embodiments of any of the compositions described herein, the β -casein protein is a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth β -casein protein.

In some embodiments of any of the compositions described herein, the composition further comprises: an alpha-lactalbumin protein at a final concentration of about 0.4g/L wt% to about 2.5 wt%; and/or a final concentration of about 2.5 wt.% to about 4.5 wt.% of beta-lactoglobulin protein. In some embodiments of any of the methods described herein, the alpha-lactalbumin protein is a bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth alpha-lactalbumin protein. In some embodiments of any of the compositions described herein, the β -lactoglobulin protein is a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth β -lactoglobulin protein.

In some embodiments of any of the compositions described herein, the composition further comprises: α -S1-casein protein at a final concentration of about 11% to about 16% by weight; and/or alpha-S2-casein protein at a final concentration of about 2 wt% to about 5 wt%. In some embodiments of any of the compositions described herein, the α -S1-casein protein is a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american caprine (mountain goat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth α -S1-casein protein; and/or the alpha-S2-casein protein is a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth alpha-S2-casein protein.

In some embodiments of any of the compositions described herein, the composition further comprises one or more of the following: serum albumin, lactoferrin and transferrin. In some embodiments of any of the compositions described herein, the serum albumin is bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly echis serum albumin; the lactoferrin is bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna or mammoth lactoferrin; and/or the transferrin is a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth transferrin.

Some embodiments of any of the compositions described herein further comprise one or more color balancing agents. In some embodiments of any of the compositions described herein, the one or more color balancing agents is beta-carotene or annatto. In some embodiments of any of the compositions described herein, the composition has a pH of about 6.2 to about 7.2 (e.g., about 6.2 to about 6.8).

Also provided are compositions comprising: mammalian-produced milk or processed mammalian-produced milk; and one or both of a kappa-casein protein that is unglycosylated or has a non-mammalian glycosylation pattern and a beta-casein protein that is unglycosylated or has a non-mammalian glycosylation pattern. In some embodiments of these methods, the final concentration of non-glycosylated or non-mammalian glycosylation pattern kappa-casein protein in the composition is from 0.02 wt.% to about 3.0 wt.%. In some embodiments of these methods, the final concentration of non-glycosylated or non-mammalian glycosylation pattern beta-casein protein in the composition is from 0.02 wt.% to about 3.0 wt.%. In some embodiments of these methods, the final concentration of non-glycosylated and/or non-mammalian glycosylation pattern kappa-casein protein in the composition is from about 0.02 wt.% to about 0.6 wt.%; and the final concentration of non-glycosylated and/or non-mammalian glycosylation pattern beta-casein in the composition is from about 0.02 wt% to about 2.5 wt%.

Also provided is a powder composition comprising: kappa-casein protein at a final concentration of about 3.6% to about 5.4% by weight; a final concentration of about 16.3% to about 24.5% by weight of beta-casein protein; a sweetener at a final concentration of about 35% to about 40% by weight; one or more lipids at a final concentration of about 25% to about 30% by weight; ash at a final concentration of about 5 wt% to about 7 wt%; and water at a final concentration of about 2 wt% to about 5 wt%, wherein the kappa-casein protein is non-glycosylated and/or has a non-mammalian glycosylation pattern, and/or the beta-casein protein is non-glycosylated and/or has a non-mammalian glycosylation pattern.

Also provided are nucleic acids comprising: a promoter; a sequence of encoded signal sequences; a sequence encoding a milk protein; and a yeast termination sequence, wherein the promoter is operably linked to a signal sequence operably linked to a sequence encoding a milk protein and the termination sequence is operably linked to a sequence encoding a milk protein. In some embodiments of these nucleic acids, the promoter is a constitutive promoter. In some embodiments of these nucleic acids, the promoter is an inducible promoter. In some embodiments of these nucleic acids, the signal sequence is a signal sequence from the encoded milk protein or a different milk protein, or a signal sequence from a yeast mating factor. In some embodiments of these nucleic acids, the encoded milk protein is selected from the group consisting of: beta-casein, kappa-casein, alpha-S1-casein, alpha-S2-casein, alpha-lactalbumin, beta-lactoglobulin, lactoferrin or transferrin. In some embodiments of these nucleic acids, the nucleic acid comprises a bacterial origin of replication. In some embodiments of these nucleic acids, the nucleic acid further comprises a selectable marker. In some embodiments of these nucleic acids, the selectable marker is an antibiotic resistance gene.

Some embodiments of these nucleic acids further comprise: an additional promoter sequence; a further sequence of encoded signal sequences; a sequence encoding an additional milk protein; and an additional yeast termination sequence, wherein the additional promoter sequence is operably linked to the additional sequence encoding the signal sequence, the sequence encoding the signal sequence is operably linked to the sequence encoding the additional milk protein, and the sequence encoding the additional milk protein is operably linked to the additional yeast termination sequence.

Also provided are host cells comprising any of the nucleic acids described herein. In some embodiments of these host cells, the host cell is a yeast strain (e.g., Kluyveromyces species, Pichia species, Saccharomyces species, Tetrahymena species, yarrowia species, Hansenula species, Brucella species: (B. sup.))Blastobotrys sp.), Candida species, Zygosaccharomyces species, or Debaryomyces species).

Also provided herein is a method of producing a recombinant milk protein that is non-glycosylated or has a non-mammalian glycosylation pattern, the method comprising: culturing any of the host cells described herein in a culture medium under conditions sufficient to allow secretion of milk proteins that are non-glycosylated or have a non-mammalian glycosylation pattern; and harvesting the milk protein that is not glycosylated or has a non-mammalian glycosylation pattern from the culture medium.

Also provided is a method of producing micelles comprising β -casein, either unglycosylated or having a non-mammalian glycosylation pattern, and κ -casein, either unglycosylated or having a non-mammalian glycosylation pattern, comprising: culturing any of the host cells provided herein in a culture medium under conditions sufficient to allow the release of micelles from the host cell, wherein the host cell comprises a nucleic acid comprising a sequence encoding beta-casein and a sequence encoding kappa-casein.

Also provided is a method of supplementing milk produced by a mammal, comprising: providing a mammal-produced milk or a processed mammal-produced milk; and mixing into the milk at least one of: a β -casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern; kappa-casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern; and micelles comprising β -casein protein that is unglycosylated or has a non-mammalian glycosylation pattern and κ -casein protein that is unglycosylated or has a non-mammalian glycosylation pattern.

Also provided is a method of producing a composition comprising: sonicating a liquid comprising beta-casein protein and kappa-casein protein or a protein mixture comprising micelles comprising beta-casein protein and kappa-casein protein; mixing the ash into a liquid; adding a mixture of one or more lipids, one or more flavor compounds, and one or more color balancing agents to the liquid, and sonicating the liquid; and adding one or more sweeteners to the liquid, thereby producing a composition. In some embodiments of these methods, the beta-casein protein is non-glycosylated or has a non-mammalian glycosylation pattern, and/or the kappa-casein protein is non-glycosylated or has a non-mammalian glycosylation pattern. In some embodiments of these methods, the ash comprises one or more of: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some embodiments of these methods, the added ash comprises CaCl ₂、KH₂PO₄And sodium citrate (e.g., one, two, or three). In some embodiments of these methods, the one or more lipids comprise at least one of: sunflower oil, coconut oil, tributyrin, mono-and diglycerides, free fatty acids and phospholipids. In some embodiments of these methods, the free fatty acid comprises at least one fatty acid selected from the group consisting of: butyric acid, caproic acid, caprylic acid and capric acid. In some embodiments of these methods, the phospholipid is soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. In some embodiments of these methods, the monoglycerides and diglycerides are vegetable-derived monoglycerides and diglycerides or are bacterial-derived monoglycerides and diglycerides. In some embodiments of these methods, the flavor compounds comprise at least one flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone. In some embodiments of these methods, the one or more color balancing agents is a beta-fiduciary Carotene or annatto. In some embodiments of these methods, the one or more sweeteners are sugars. In some embodiments of these methods, the saccharide is selected from the group consisting of: glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. In some embodiments of these methods, the one or more sweeteners are artificial sweeteners. In some embodiments of these methods, the artificial sweetener is selected from the group consisting of: stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osthol, betadine, sorbitol, thaumatin, xylitol, potassium acetamidosulfonate, advatame, alitame, aspartame-acesulfame, cyclamate, dulcin, neohesperidin dihydrochalcone, neotame, and P-4000. In some embodiments of these methods, the pH of the liquid is from about 6.2 to about 7.4 (e.g., from about 6.4 to about 6.8). In some embodiments of these methods, the β -casein protein is a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna or woolly down-casein; and/or the kappa-casein protein is a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth kappa-casein protein. In some embodiments of these methods, the protein mixture further comprises one or more of A plurality of proteins selected from the group consisting of: alpha-lactalbumin, beta-lactoglobulin, alpha-S1-casein, alpha-S2-casein, lactoferrin, transferrin and serum albumin.

Also provided are compositions produced by any of the methods described herein.

Also provided is a method of preparing butter, cheese, caseinate or yoghurt comprising: providing any of the compositions described herein; and producing butter, cheese, caseinate or yoghurt using any of the compositions described herein as starting material.

Also provided is a kit comprising: (a) a mixture of one or more milk proteins, one or more fats and one or flavor compounds; and (b) a mixture of ash and at least one sweetener. In some embodiments of these kits, the one or more milk proteins are selected from the group consisting of: beta-casein, kappa-casein, alpha-lactalbumin, beta-lactoglobulin, alpha-S1-casein, alpha-S2-casein, lactoferrin, transferrin and serum albumin. In some embodiments of these kits, the one or more milk proteins are cow, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, marsupial (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, gill, or woolly mammoth milk proteins. In some embodiments of these kits, the one or more fats are selected from: sunflower oil, coconut oil, tributyrin, mono-and diglycerides, free fatty acids and phospholipids. In some embodiments of these kits, the free fatty acid comprises at least one fatty acid selected from the group consisting of: butyric acid, caproic acid, caprylic acid and capric acid. In some embodiments of these kits, the phospholipid is soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. In some embodiments of these kits, the monoglycerides and diglycerides are plant-derived monoglycerides Glycerides and diglycerides, or monoglycerides and diglycerides of bacterial origin. In some embodiments of these kits, the flavor compounds comprise at least one flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone. In some embodiments of these kits, the mixture in (a) further comprises one or more color balancing agents. In some embodiments of these kits, the one or more color balancing agents is beta-carotene or annatto. In some embodiments of these kits, the one or more sweeteners are sugars (e.g., sugars selected from the group consisting of glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose). In some embodiments of these kits, the one or more sweeteners are artificial sweeteners (e.g., an artificial sweetener selected from the group consisting of stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osthol, betadine, sorbitol, thaumatin, xylitol, potassium acetamido sulfonate, avantam, alitame, aspartame-acesulfame salt, sodium cyclamate, dulcin, neohesperidin dihydrochalcone, neotame, and P-4000). In some embodiments of any of these kits, the ash comprises one or more of: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some embodiments of these kits, the ash comprises CaCl ₂、KH₂PO₄And sodium citrate (e.g., one, two, or three). Some embodiments of these kits further comprise instructions for preparing any of the compositions described herein.

Kits comprising at least one nucleic acid described herein are also provided.

Also provided herein is a dairy substitute food product comprisingOne or more of an isolated milk protein component, fat, carbohydrate, and ash. In some embodiments of these dairy substitute food products, the food product is of non-animal origin. In some embodiments of these alternative food products, the food product comprises milk, butter, cheese, caseinate, yogurt, and cream. In some embodiments of these dairy replacement foods, the isolated milk protein component comprises casein and whey protein. In some embodiments of these dairy replacement foods, the casein proteins further include α -s1, α -s2, β, and κ -casein. In some embodiments of these dairy replacement foods, the casein proteins also include alpha-s 1, beta, and kappa-casein proteins. In some embodiments of these dairy substitute foods, the casein protein further comprises a component for micelle formation. In some embodiments of these dairy substitute foods, the casein protein exhibits milk clotting properties at a pH of 4.0-6.0. In some embodiments of these dairy substitute foods, the casein protein is at least or equal to 2.5% (w/v) and less than or equal to 10% (w/v). In some embodiments of these dairy replacement foods, the whey protein further comprises beta-lactoglobulin and alpha-lactalbumin. In some embodiments of these dairy replacement foods, the whey protein forms a polymer matrix gel. In some embodiments of these dairy replacement foods, the whey protein is at least 0.1% (w/v) and less than or equal to 1% (w/v). In some embodiments of these dairy substitute foods, the one or more milk protein components are separated from the microorganisms. In some embodiments of any of these dairy substitute foods, the one or more milk protein components are isolated from the recombinant microorganism. In some embodiments of these dairy substitute foods, the one or more milk protein components are synthesized in eukaryotic microorganisms. In some embodiments of these dairy substitute foods, the eukaryotic microorganism comprises yeast. In some embodiments of these dairy substitute foods, the yeast includes Kluyveromyces species, Pichia species, Saccharomyces species, and Tetrahymena species (b: (b)) Tetrahymena sp.)。

In some embodiments of these alternative foods, the fat comprises triglycerides. In some embodiments of these dairy replacement foods, the fat comprises a high oleic oil. In some embodiments of these dairy replacement foods, the high oleic oil further comprises one or more of monounsaturates, oleic acid, linoleic acid, linolenic acid, and saturates. In some embodiments of these dairy substitute foods, the fat comprises a short chain fatty acid. In some embodiments of these dairy replacement foods, the short chain fatty acids include butyric acid, caproic acid, caprylic acid, and capric acid. In some embodiments of these dairy replacement foods, the one or more fats comprise trans-esterified fatty acids. In some embodiments of these dairy substitute foods, the one or more fats are isolated from a plant. In some embodiments of these dairy substitute foods, the plant is selected from one or more of the following: sunflower, corn, olive, soybean, peanut, walnut, almond, sesame, cottonseed, canola, safflower, linseed, palm kernel, palm fruit, coconut, babassu, shea butter, mango butter, cocoa butter, wheat germ, and rice bran oil. In some embodiments of these dairy substitute foods, the sugars comprise galactose, sucrose, glucose, fructose, and maltose. In some embodiments of these dairy replacement foods, the dairy replacement food is substantially free of lactose. In some embodiments of these dairy substitute foods, the ash comprises minerals. In some embodiments of these dairy substitute foods, the minerals further comprise one or more of the following: sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc, chloride, manganese, selenium, iodine, retinol, carotene, vitamins, vitamin D, vitamin E, vitamin B12, thiamine, and riboflavin. In some embodiments of these dairy substitute foods, the ash comprises an anion. In some embodiments of these dairy substitute foods, the minerals further comprise one or more of the following: phosphates, citrates, sulfates, carbonates and chlorides.

Also provided is a method of making a dairy substitute food product comprising the step of contacting one or more of an isolated milk protein component, an interesterified fat, a carbohydrate, and an ash. Some embodiments of these methods further comprise the step of isolating one or more milk protein fractions from the lower eukaryote.

Also provided is a method of modifying the flavor profile of a dairy substitute product comprising modulating the combination of fatty acids in a mixture comprising a milk protein component, a carbohydrate, and ash. In some embodiments of these methods, the adjusting step comprises triglycerides comprising three oleic acids and short chain triglycerides comprising butyric acid, a caproic acid and a caprylic acid. In some embodiments of these methods, the modulating step comprises increasing or decreasing fatty acids comprising butyric acid, caproic acid, caprylic acid, and capric acid. In some embodiments of these methods, the flavor profile of the dairy substitute product mimics the flavor profile of one or more dairy products. In some embodiments of these methods, the flavor profile of the one or more dairy products comprises cow's milk, goat's milk, soy milk, almond milk, and coconut milk. In some embodiments of these methods, the flavor profile comprises one or more sensory impressions selected from the group consisting of: buttery, nutty, sweet, sour, fruity, floral, bitter, woody, earthy, beany, spicy, metallic, sweet, musty, oily and vinegar flavored.

Disclosed herein are methods and compositions for producing a dairy substitute product. In some embodiments, methods and compositions are provided for dairy replacement foods comprising one or more isolated milk protein components, fat, carbohydrates, and ash. In certain embodiments, methods and compositions are provided for a dairy replacement composition comprising casein protein and whey protein, wherein the composition is substantially free of animal products, and wherein the ratio of casein protein to whey protein is preferably a preferred (w/v) ratio. In certain other embodiments, methods of modulating the flavor profile of a dairy substitute food product are provided that include modulating the fatty acid content in a mixture comprising a milk protein component, fat, carbohydrate, and ash. Preferred conditioning steps include increasing or decreasing one or more fatty acids comprising butyric acid, caproic acid, caprylic acid and capric acid. In further embodiments, the methods and compositions of the present invention provide a desired (w/v) ratio of milk protein component and fat.

In various aspects, the methods and compositions of the present invention provide a dairy substitute that still retains its functional characteristics and organoleptic properties. In some embodiments, the core function may be, but is not limited to, achieving nutritional characteristics similar to conventional dairy products, and replicating one or more (if not all) of its core functions.

In other embodiments, the core function may be, but is not limited to, replicating the same or similar sensory characteristics as traditional dairy-based products, including, but not limited to, taste, appearance, handling and mouthfeel, desired density, structure, texture, resilience, elasticity, coagulation, binding, fermentation, aeration, foaming, creaminess, and emulsification.

Preferred methods and compositions provide dairy product replacement products such as milk, butter, cheese, yogurt and cream. Provided herein are formulations of non-dairy milk replacers comprising (3.3%) one or more isolated milk protein components, (4.0%) fat, (2.4%) carbohydrates and (0.7%) ash (w/v). The flavor profile of the non-dairy milk substitute is enhanced by altering the fat content by adjusting the triglyceride level and the fatty acid composition of the triglycerides.

Advantages of the methods and dairy substitute compositions include reduction or removal of antibiotic residues, heavy metals, bacteria and adulterants commonly found in natural dairy products and reduction of environmental impact.

Accordingly, certain aspects of the present invention provide an animal component-free dairy substitute that has desirable flavor characteristics, such as replicating dairy flavor, minimizing food-borne pathogens, and having a lower environmental impact, while maintaining downstream applications of dairy milk.

Unless defined otherwise herein, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by one of ordinary skill in the art. Furthermore, unless the context requires otherwise, singular terms shall include plural terms and plural terms shall include singular terms. Generally, the nomenclature used in connection with, and the techniques of, dairy processing, biochemistry, enzymology, molecular and cellular biology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art.

Methods and materials for use in the present invention are described herein; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and not intended to be limiting.

All publications, patents, patent applications, sequences, database entries, and other references mentioned herein are incorporated by reference to the same extent as if each individual publication, patent application, sequence, database entry, and other reference were specifically and individually indicated to be incorporated by reference. In case of conflict, the present specification, including definitions, will control.

Other features and advantages of the invention will be apparent from the following detailed description and drawings, and from the claims.

The terms and descriptions used herein are used for the purpose of describing particular embodiments only and are not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. The terms "comprising," including, "" having, "" with, "or variants thereof, are intended to be inclusive in a manner similar to the term" comprising.

An "isolated" RNA, DNA, or conjunct polymer is one that is substantially separated from other cellular components that naturally accompany the native polynucleotide in its native host cell (e.g., ribosomes, polymerases, and genomic sequences with which it is naturally associated).

As used herein, an "isolated" organic molecule (e.g., fatty acid or SCFA) is an organic molecule that is substantially separated from the cellular components (membrane lipids, chromosomes, proteins) of the host cell from which it originates. As used herein, the term "isolated" with respect to a protein means that the preparation of the protein is at least 60% pure, e.g., greater than 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% pure. The term does not require that the biomolecule has been separated from all other chemicals, although some of the separated biomolecules may be purified to near homogeneity.

The term "polynucleotide" or "nucleic acid molecule" refers to a polymeric form of nucleotides that are at least 10 bases in length. The term includes DNA molecules (e.g., cDNA or genomic or synthetic DNA) and RNA molecules (e.g., mRNA or synthetic RNA), as well as analogs of the DNA or RNA that contain non-natural nucleotide analogs, non-natural internucleoside linkages, or both. The nucleic acid may be in any topological conformation. For example, the nucleic acid may be single-stranded or double-stranded or circular.

The term "SCFA" is an abbreviation for short chain fatty acids, the term "HOSO" is an abbreviation for high oleic sunflower oil, and "SCTG" is an abbreviation for short chain triglycerides.

The term "milk protein component" refers to proteins or protein equivalents and variants found in milk, such as casein, whey, or combinations of casein and whey, including subunits thereof, derived from various sources and as further defined herein.

The term "milk protein" refers to a protein found in, or having a sequence that is at least 80% identical (e.g., at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to a sequence of a protein found in, milk produced by a mammal. Non-limiting examples of milk proteins include: beta-casein, kappa-casein, alpha-S1-casein, alpha-S2-casein, alpha-lactalbumin, beta-lactoglobulin, lactoferrin, transferrin and serum albumin. Additional milk proteins are known in the art.

The term "casein protein" is known in the art and denotes a family of proteins that are present in mammalian produced milk and are capable of self-assembling with other proteins in the family to form micelles and/or precipitate out of an aqueous solution at acidic pH. Non-limiting examples of casein proteins include: beta-casein, kappa-casein, alpha-S1-casein and alpha-S2-casein. Provided herein are non-limiting examples of sequences of casein proteins from different mammals. Additional sequences of other mammalian caseins are known in the art.

The term "mammal-produced milk" is known in the art and refers to milk produced by a mammal.

The term "processed mammal produced milk" refers to mammal produced milk that is processed using one or more steps known in the dairy industry (e.g., homogenization, pasteurization, irradiation, or supplementation).

The term "component of mammalian origin" refers to a molecule or compound (e.g., a protein, lipid, or nucleic acid) obtained from the body of a mammal or a molecule obtained from a fluid or solid produced by a mammal.

The term "component of milk" or "milk component" is a molecule, compound, element or ion present in milk produced by a mammal.

The term "non-mammalian glycosylation pattern" refers to one of a difference in one or more positions of glycosylation in a protein produced and post-translationally modified in a non-mammalian cell (e.g., a yeast cell, an insect cell, or a bacterial cell) and/or a difference in the amount and/or type of glycosylation at one or more positions in the protein as compared to a reference protein (e.g., the same protein produced and post-translationally modified in a mammalian cell, such as a CHO cell, a MEK cell, or a mammalian mammary cell)).

The term "lipid" refers to one or more molecules (e.g., biomolecules) that include a fatty acyl group (e.g., a saturated or unsaturated acyl chain). For example, the term lipid includes oils, phospholipids, free fatty acids, phospholipids, monoglycerides, diglycerides, and triglycerides. Non-limiting examples of lipids are described herein. Additional examples of lipids are known in the art.

The term "plant-derived lipid" refers to a lipid obtained and/or produced from a plant (e.g., a monocot or dicot).

The term "sweetener" refers to a sugar (e.g., a monosaccharide, disaccharide, or polysaccharide) or an artificial sweetener (e.g., a small molecule artificial sweetener or a protein artificial sweetener) that when added to a composition causes the composition to taste sweet when ingested by a mammal, such as a human. Non-limiting examples of sweeteners are described herein. Additional examples of sweeteners are known in the art.

The term "ash" is a term known in the art and refers to one or more ions, elements, minerals and/or compounds that may be found in milk produced by a mammal. Non-limiting ions, elements, minerals, and compounds found in mammalian produced milk are described herein. Additional ions, elements, minerals and compounds found in mammalian produced milk are also known in the art.

The term "color balancing agent" or "colorant" refers to an agent added to a composition to adjust the color of the composition, for example, to make the color of the composition look more similar to milk produced by a mammal. Non-limiting examples of color balancing agents or colorants include beta-carotene and annatto. Other examples of color balancing agents are known in the art. The color balancing agent or colorant may be produced or obtained from a plant.

The term "micelle" refers to a generally (or approximately) spherical supramolecular structure that exists as a dispersion in a composition. The micelle may have a surface consisting of, for example, a charged outer layer. Micelles may encapsulate one or more biomolecules. For example, micelles may encapsulate two or more proteins (e.g., β -casein protein and κ -casein protein). The micelle may have a diameter of about 10nm to about 350 nm. Additional aspects and features of micelles are known in the art.

The phrase "concentration of a component in milk produced by a mammal" refers to the concentration of a component in milk produced by a mammal or the average concentration of a component in milk produced by a population of mammals of the same species.

The term "attenuation" as used herein generally refers to a loss of function, including mutation, partial or complete deletion, insertion or other modification, of a gene sequence or a sequence controlling transcription of a gene sequence that reduces or inhibits production of a gene product, or renders a gene product non-functional. In some cases, the loss of function is described as a knockout mutation. Attenuation also includes amino acid sequence changes by altering the nucleic acid sequence, placing a gene under the control of a less active promoter, down-regulating, expressing interfering RNA, ribozymes, or antisense sequences that target the gene of interest, or by any other technique known in the art. In one example, the sensitivity of a particular enzyme to feedback inhibition or inhibition caused by a composition that is not a product or reactant (non-pathway-specific feedback) is reduced such that the enzyme activity is not affected by the presence of the compound. In other cases, an enzyme that has been altered to a lower activity may be referred to as attenuated.

Deletion (c): one or more nucleotides are removed from the nucleic acid molecule or one or more amino acids are removed from the protein, and the regions on either side are ligated together.

Knocking out: a gene whose expression or activity level has been reduced to zero. In some examples, a gene is knocked out via deletion of some or all of its coding sequence. In other examples, a gene is knocked out by introducing one or more nucleotides into its open reading frame, which results in translation of a nonsense or otherwise non-functional protein product.

The term "synthetic milk substitute" refers to a composition that is similar, analogous, equal, or nearly identical to dairy milk.

The term "flavor" refers to the taste and/or aroma of a food or beverage.

The term "recombinant" is a term known in the art. The term "recombinant" when referring to a nucleic acid (e.g., a gene) can be used, for example, to describe a nucleic acid that has been removed from its naturally occurring environment, a nucleic acid that is not associated with all or a portion of the nucleic acid adjacent or proximal to the nucleic acid when it is found in nature, a nucleic acid that is operably linked to a nucleic acid to which it is not linked in nature, or a nucleic acid that does not occur in nature. The term "recombinant" may be used, for example, to describe a cloned DNA isolate, or a nucleic acid comprising a chemically synthesized nucleotide analog. When "recombinant" is used to describe a protein, it may refer to, for example, a protein produced in a different species or type of cell than the species or type of cell that produces the protein in nature.

As used herein, an endogenous nucleic acid sequence (or the encoded protein product of that sequence) in the genome of an organism is considered "recombinant" herein if a heterologous sequence is placed adjacent to the endogenous nucleic acid sequence such that expression of the endogenous nucleic acid sequence is altered. In this context, a heterologous sequence is a sequence that is not naturally adjacent to an endogenous nucleic acid sequence, whether the heterologous sequence itself is endogenous (derived from the same host cell or progeny thereof) or exogenous (derived from a different host cell or progeny thereof). For example, a promoter sequence can replace (e.g., by homologous recombination) a native promoter of a gene in the genome of a host cell such that the gene has an altered expression pattern. The gene will now become "recombinant" in that it is separated from at least some of the sequences that naturally flank it.

A nucleic acid is also considered "recombinant" if it contains any modifications that do not naturally occur to the corresponding nucleic acid in the genome. For example, an endogenous coding sequence is considered "recombinant" if it contains an insertion, deletion, or point mutation introduced artificially (e.g., by human intervention). "recombinant nucleic acid" also includes nucleic acids that integrate into the host cell chromosome at a heterologous site and nucleic acid constructs that exist as episomes.

The term "percent sequence identity" or "identity" in the context of nucleic acid sequences refers to the residues in the two sequences that are the same when aligned for maximum correspondence. The length of the sequence identity comparison may be at least about nine nucleotides, typically at least about 20 nucleotides, more typically at least about 24 nucleotides, typically at least about 28 nucleotides, more typically at least about 32 nucleotides, and preferably at least about 36 orA segment of more nucleotides. There are many different algorithms known in the art that can be used to measure nucleotide sequence identity. For example, the polynucleotide sequences can be compared using FASTA, Gap or Bestfit, a program in Wisconsin Package Version 10.0, Genetics Computer Group (GCG), Madison, Wis. FASTA provides alignments and percent sequence identities of the best overlapping regions between query and search sequences. See, for example, Pearson,Methods Enzymol.183:63-98, 1990 (which is incorporated herein by reference in its entirety). For example, percent sequence identity between nucleic acid sequences can be determined using FASTA with its default parameters (word length 6, NOPAM factor of the scoring matrix) or using Gap with its default parameters as provided in GCG version 6.1, which is incorporated herein by reference. Alternatively, the computer program BLAST (Altschul et al, J. Mol. Biol.215:403-,Nature Genet.3:266-,Meth. Enzymol.266:131, 141, 1996; Altschul et al,Nucleic Acids Res.25:3389-3402, 1997, Zhang and Madden,Genome Res.649-656, 1997), in particular blastp or tblastn (Altschul et al,Nucleic Acids Res.25:3389-3402, 1997).

The term "substantial homology" or "substantial similarity", when referring to a nucleic acid or fragment thereof, means that there is nucleotide sequence identity in at least about 76%, 80%, 85%, preferably at least about 90%, more preferably at least about 95%, 96%, 97%, 98% or 99% of the nucleotide bases, when optimally aligned with an appropriate nucleotide insertion or deletion with another nucleic acid (or its complementary strand), as measured by any well-known sequence identity algorithm, such as FASTA, BLAST or Gap as discussed above.

Alternatively, substantial homology or similarity exists when a nucleic acid or fragment thereof hybridizes under stringent hybridization conditions to another nucleic acid, a strand of another nucleic acid, or a complementary strand thereof. In the context of nucleic acid hybridization experiments, "stringent hybridization conditions" and "stringent wash conditions" depend on a number of different physical parameters. Nucleic acid hybridization will be affected by conditions such as salt concentration, temperature, solvent, base composition of the hybridizing species, length of the complementary region, and the number of nucleotide base mismatches between hybridizing nucleic acids, as will be readily understood by those skilled in the art. One of ordinary skill in the art would know how to vary these parameters to achieve a particular stringency of hybridization.

In general, under a particular set of conditions, a "stringent hybridization" is performed at about 25 ℃ at the thermal melting point (Tm) for a particular DNA hybrid. Under a particular set of conditions, a "stringent wash" is performed at a temperature of about 5 ℃ at the Tm of a particular DNA hybrid. The Tm is the temperature at which 50% of the target sequence hybridizes to a perfectly matched probe. See Sambrook et al, Molecular Cloning, A Laboratory Manual, 2 nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., page 9.51, 1989, which is incorporated herein by reference. For purposes herein, "stringent conditions" are defined as aqueous hybridization (i.e., without formamide) in 6XSSC (where 20XSSC contains 3.0M NaCl and 0.3M sodium citrate), 1% SDS at 65 ℃ for 8-12 hours followed by two washes in 0.2XSSC, 0.1% SDS at 65 ℃ for 20 minutes for solution-phase hybridization. One skilled in the art will appreciate that hybridization at 65 ℃ will occur at different rates, depending on a number of factors, including the length and percent identity of the hybridizing sequences.

Nucleic acids (also referred to as polynucleotides) of the invention may include RNA, cDNA, genomic DNA, and sense and antisense strands in synthetic form, as well as mixed polymers of the foregoing. They may be chemically or biochemically modified, or may contain non-natural or derivatized nucleotide bases, as will be readily understood by those skilled in the art. Such modifications include, for example, labels, methylation, substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoramidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), overhangs (e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.), chelators, alkylating agents, and modified linkages (e.g., alpha anomeric nucleic acids, etc.). Examples of modified nucleotides are described in Malyshiev et al, Nature509,385-,J. Am. Chem. Soc.136:826-829, 2014. Also included are synthetic molecules that mimic polynucleotides in their ability to bind a designated sequence via hydrogen bonding and other chemical interactions. Such molecules are known in the art and include, for example, those in which peptide chains replace the phosphate ester linkages in the backbone of the molecule. Other modifications may include, for example, analogs in which the ribose ring contains a bridging moiety or other structure such as those found in "locked" nucleic acids.

The term "mutated" when applied to a nucleic acid sequence means that a nucleotide in the nucleic acid sequence may be inserted, deleted or altered as compared to a reference nucleic acid sequence. A single alteration (point mutation) may be made at a locus, or multiple nucleotides may be inserted, deleted or altered at a single locus. In addition, one or more changes may be made at any number of loci within a nucleic acid sequence. Nucleic acid sequences can be mutated by any method known in the art, including, but not limited to, mutagenesis techniques such as "error-prone PCR" (a method in which PCR is performed under conditions in which the replication fidelity of the DNA polymerase is low, such that a high point mutation rate is obtained along the entire length of the PCR product; see, e.g., Leung et al, Technique1:11-15, 1989, and Caldwell and Joyce,PCR Methods Applic.2:28-33, 1992); and "oligonucleotide-directed mutagenesis" (a method capable of generating site-specific mutations in the DNA fragment of any clone of interest; see, e.g., Reidhaar-Olson and Sauer,Science 241:53-57, 1988)。

the term "vector" as used herein means a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid," which generally refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated, but also includes linear double-stranded molecules, such as those resulting from amplification by Polymerase Chain Reaction (PCR) or treatment of the circular plasmid with a restriction enzyme. Other vectors include cosmids, Bacterial Artificial Chromosomes (BACs) and Yeast Artificial Chromosomes (YACs). Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome (discussed in more detail below). Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., vectors having an origin of replication that functions in the host cell). Other vectors may be introduced into a host cell and integrated into the genome of the host cell, and thereby replicated together with the host genome. In addition, certain preferred vectors are capable of directing the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply "expression vectors").

Promoters useful for expression of the recombinant genes described herein include constitutive and inducible/repressible promoters. Examples of inducible/repressible promoters include galactose-inducible promoters (e.g., PLAC 4-PBI). When multiple recombinant genes are expressed in engineered yeast, different genes may be controlled by different promoters or by different promoters in different operons, or expression of two or more genes may be controlled by a single promoter that is part of an operon.

The term "operably linked" expression control sequence refers to a bond wherein the expression control sequence is contiguous with a gene of interest to control the gene of interest and with an expression control sequence that functions in trans or at a distance to control the gene of interest.

The terms "expression control sequence" or "regulatory sequence" are used interchangeably and, as used herein, refer to a polynucleotide sequence necessary to effect expression of a coding sequence to which it is operably linked. Expression control sequences are sequences that control the transcription, post-transcriptional events, and translation of a nucleic acid sequence. Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; effective RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that increase translation efficiency (e.g., ribosome binding sites); sequences that enhance protein stability; and, when desired, sequences that enhance protein secretion. The nature of such control sequences varies depending on the host organism; in prokaryotes, such control sequences typically include a promoter, a ribosome binding site, and a transcription termination sequence. The term "control sequences" is intended to include at least all components whose presence is essential for expression, and may also include additional components whose presence is advantageous, such as leader sequences and fusion partner sequences.

The terms "transfection", "transfection" and the like refer to the introduction of heterologous nucleic acid into eukaryotic cells (higher and lower eukaryotic cells). Historically, the term "transformation" has been used to describe the introduction of nucleic acids into yeast or fungal cells; however, in this context, the term "transfection" is used to refer to the introduction of nucleic acids into any eukaryotic cell, including yeast and fungal cells.

The term "recombinant host cell" ("expression host cell", "expression host system", "expression system" or simply "host cell") as used herein means a cell into which a recombinant vector has been introduced. It is understood that these terms refer not only to the particular subject cell, but also to the progeny of such a cell. Because certain modifications may occur in the progeny due to mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the host cell as used herein. The recombinant host cell may be an isolated cell or cell line grown in culture, or may be a cell that resides in a living tissue or organism. Preferred host cells are yeasts and fungi.

The terms "yeast and filamentous fungi" include, but are not limited to, any Kluyveromyces such as Kluyveromyces lactis, Kluyveromyces marxianus, Saccharomyces such as Saccharomyces cerevisiae, Pichia such as Pichia pastoris, Pichia finnishis, Pichia trehalose, Pichia pastoris, Pichia membranaefaciens, Pichia pastoris: (A)Ogataea minutaPichia linderae)opuntiaePichia pastoris, heat-resistant Pichia pastoris,salictariaPichia pastoris,guercuumPichia pastoris, Pichia pickettii,stiptisPichia pastoris, Pichia methanolica, Hansenula polymorpha, Candida albicans, any Aspergillus genus, such as Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium, FusariumBelonging to the genera Fusarium graminearum, Fusarium, Physcomitrella patens and Neurospora crassa.

As used herein, the term "predominantly" or variants thereof will be understood to refer to, for example, a) the amount of a particular fatty acid composition relative to the total amount of fatty acid composition in the context of fat; b) in the context of proteins, the amount of a particular protein composition (e.g., beta-casein) relative to the total amount of the protein composition (e.g., alpha-, beta-, and kappa-casein).

The terms "about," "approximately," or "similar to" mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, or the limitations of the measurement system. It is to be understood that all ranges and amounts described below are approximate and are not intended to limit the invention. Where ranges and numbers are used, these may be approximate to include statistical ranges or measurement errors or variations. In some embodiments, for example, the measurement may be plus or minus 10%.

The phrase "substantially free of" is used to indicate that the indicated component, if present, is present in an amount that does not contribute, or contributes only in a minimal manner, to the properties of the composition. In various embodiments, where a composition is substantially free of a particular component, the component is present in less than a functional amount. In various embodiments, the components may be present in trace amounts. The particular limit will vary depending on the nature of the components, but may be, for example, selected from less than 10 wt%, less than 9 wt%, less than 8 wt%, less than 7 wt%, less than 6 wt%, less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt%, less than 1 wt%, or less than 0.5 wt%.

As used herein, the term "substantially free" of a particular carbohydrate, such as lactose, is used to indicate that the food composition is substantially free of carbohydrate residues. By purity, substantially free means that the amount of carbohydrate residues does not exceed 10%, preferably below 5%, more preferably below 1%, most preferably below 0.5%, wherein the percentages are by weight or by mole. Thus, substantially all carbohydrate residues in the food composition according to the invention are free of e.g. lactose.

Unless otherwise indicated, percentages (%) of ingredients refer to total weight%.

Unless otherwise indicated, and as an example of all sequences described under the general format "SEQ ID NO:", a "nucleic acid comprising SEQ ID NO: 1" refers to a nucleic acid, at least a portion of which has (i) the sequence of SEQ ID NO:1, or (ii) a sequence complementary to SEQ ID NO: 1. The choice between the two is determined by the context. For example, if a nucleic acid is used as a probe, the choice between the two is determined by the requirement that the probe be complementary to the desired target.

Brief Description of Drawings

Figure 1 shows a flow diagram representing an exemplary method of producing a synthetic milk substitute.

Fig. 2A shows a picture depicting a precipitate of an exemplary milk protein component.

Fig. 2B shows a picture depicting exemplary aggregates of milk protein components.

FIG. 3 shows an image of a silver stained SDS-PAGE gel visualizing the protein components of milk.

FIG. 4 is a SYPRO Ruby stained SDS-PAGE gel showing the level of α -lactalbumin secretion mediated by the OST signal sequence, the native α -lactalbumin signal sequence and the α mating factor signal sequence, as described in example 6.

Figure 5 shows the levels of α -lactalbumin secretion (determined by ELISA assay as described in example 6) by wild-type yeast cells using the native α -lactalbumin signal peptide or OST1 signal peptide, or by yeast cells expressing α -lactalbumin.

FIG. 6 shows levels of beta-lactoglobulin secretion (using SDS-PAGE) by wild-type yeast cells and yeast cells including vectors as described in example 6.

FIG. 7 is a Western blot showing levels of beta-lactoglobulin secreted by wild type yeast and yeast cells comprising the vector as described in example 6.

FIG. 8 is a graph showing the levels of secreted beta-casein and secreted alpha-S1-casein produced by wild type yeast and yeast cells comprising the vector as described in example 6.

Fig. 9 is a schematic diagram showing steps in the method described in example 7.

Fig. 10 is an image of a composition made by the methods described herein.

Detailed Description

The present invention is based on the discovery that the presence of only a few components in mammalian-produced milk provides the texture and taste of mammalian-produced milk, as well as the development of compositions having similar taste, aroma and mouthfeel as compared to mammalian-produced milk. In view of this discovery, provided herein are such compositions, methods of making compositions, and kits comprising these compositions and mixtures useful for making these compositions.

The compositions provided herein provide compositions having similar taste, mouthfeel, aroma, and nutritional value as compared to mammalian-produced milk, but lacking one or more components of mammalian-produced milk that may be considered undesirable (e.g., allergens, lactose, antibiotics, hormones (e.g., stress hormones and/or growth hormones), heavy metals, bacteria (e.g., escherichia coli), viruses, and prions). The compositions provided herein have improved shelf life compared to mammalian-produced milk, and may have improved flavor characteristics compared to mammalian-produced milk.

Also provided herein are methods and compositions for a dairy replacement food product comprising one or more isolated milk protein components, fat, carbohydrates, and ash. In certain aspects, the methods and compositions comprise milk or milk-like protein equivalents. Preferably, the milk protein component is substantially free of impurities. In some embodiments, the milk protein component comprises microbially-derived or produced casein, whey, or a combination thereof. More preferably, methods of introducing engineered nucleic acid sequences encoding one or more milk protein components are provided. Even more preferably, the milk protein component is not of animal origin. In other preferred embodiments, the recombinant milk protein component is modified to express the same phosphate groups or to lack phosphate groups and/or carbohydrate groups linked to casein proteins. Recombinant β -casein and recombinant κ -casein are capable of forming micelles by having the same phosphate groups of the recombinant β -casein and κ -casein as the same proteins present in milk produced by a mammal.

The methods and techniques of the present invention are generally performed according to conventional methods well known in the art, and as described in various general and more specific references that are cited and discussed throughout the present specification, unless otherwise indicated. See, for example, Sambrook et al, Molecular Cloning: A Laboratory Manual, 2 nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989, Ausubel et al, Current Protocols in Molecular Biology, Green Publishing Associates, 1992, and Supplements to 2002), Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1990, Taylor and Drickar, International to Global, Oxford Unit V.Press, 2003, Worthington Man, engineering Handbook, handb.

Exemplary materials and methods for use in any of the methods and compositions are described below, and can be used in any combination. Additional materials and methods useful in any method and composition are also known in the art.

Casein protein

Casein proteins include a number of different proteins found in mammalian milk. Non-limiting examples of casein proteins include: beta-casein, kappa-casein, alpha-S2-casein and alpha-S1-casein.

As an alternative to obtaining casein protein from a mammal or milk produced by a mammal for use in the manufacture of dairy products, the present invention provides methods and compositions for producing recombinant casein protein. In various aspects of the invention, methods and compositions are provided for non-animal derived casein proteins having similar solubility and similar turbidity, as well as thermal stability suitable for incorporation into various food products. Preferably, the casein of non-animal origin has excellent solubility, similar turbidity and heat stability suitable for incorporation into various dairy substitute products. In addition, further features of the protein include less or no aggregation or precipitation during such heat treatment and suitability for procedures such as pasteurization, concentration, and the like.

Functional differences in milk casein of non-animal origin can be characterized in the following respects: the viscosity of the liquid; the ability of a protein to withstand heat; the ability of a protein to form micelles; and the ability of proteins to retain different minerals and vitamins.

Beta-casein

Such as by Greenberg et al (J. Biol. Chem.259:5132-5138, 1984) shows that the primary structure of human beta-casein is a phosphorylated protein having phosphorylation sites at specific seryl and threonyl residues located near the amino terminus. Comparison of human and bovine beta-casein showed 47% identity.

Non-limiting examples of beta-casein proteins are SEQ ID Nos. 25, 27, 29, 31, 33, 35, 36, 38, 40, 42, 44 and 46. Non-limiting examples of nucleic acid sequences encoding beta-casein proteins are SEQ ID Nos. 26, 28, 30, 32, 34, 37, 39, 41, 43, 45, 47 and 144. The β -casein protein may be a β -casein protein from any mammalian species, for example a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, son, tiger, echidna or woolly mammoth β -casein protein. Additional sequences of different beta-casein proteins and nucleic acids encoding different beta-casein proteins are known in the art.

The beta-casein protein may also be at least 50% (e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type beta-casein protein (e.g., SEQ ID Nos: 25, 27, 29, 31, 33, 35, 36, 38, 40, 42, 44, or 46). The nucleic acid encoding a beta-casein protein may encode a protein that is at least 50% (e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type beta-casein protein (e.g., SEQ ID Nos: 25, 27, 29, 31, 33, 35, 36, 38, 40, 42, 44, or 46).

Known methods for isolating beta-casein from genetically engineered bacterial cells generally involve precipitation of beta-casein from supernatant derived from lysed or fractionated cells. For example, Simons, et al, Protein Eng.6:763, 770 (1993), using genetically engineered E.coli to express bovine beta-casein. Proteins accumulated in the periplasmic space of the bacteria are released into the cell suspension by osmotic shock. After centrifugation of the suspension, the beta-casein in the pellet was resuspended in a cold water wash and centrifuged again. The beta-casein present in the supernatant was precipitated by acidification with acetic acid, filtered and further purified by HPLC. Similarly, the flow rates of the first, Hansson, et al,Protein Express. Purif.4: 373. sup. 381, 1993, using genetically engineered E.coli for expression of beta-casein. Beta-casein present in the cell lysate was precipitated with ammonium sulfate, dissolved in ethanolamine and 6M urea, and further purified by ion exchange chromatography. See, for example, U.S. patent No. 6,121,421.

Furthermore, simpler and more efficient methods for isolating recombinantly produced beta-casein in yeast than known techniques are also known. In the case of Choi et al,J. Agric. Food Chem.49(4) 1761-Expression and purification of glycosylated bovine beta-casein (L70S/P71S) led to the following observations: most bovine beta-casein is not hyperglycosylated in pichia pastoris and its molecular weight is estimated to be 33.6 kDa. Glycosylated bovine beta-casein is typically phosphorylated to the same extent as native bovine beta-casein.

Kappa-casein

Both kappa-casein is phosphorylated and glycosylated. The sequence of human kappa-casein was determined by Brignon et al (Fed. Eur. biol. Soc. Lett. 188:48-54, 1985). See, for example, U.S. patent No. 5,710,044.

Non-limiting examples of kappa-casein proteins are SEQ ID Nos. 1, 3, 5,7, 9, 11, 13, 15, 17, 19, 21 and 23. Non-limiting examples of nucleic acid sequences encoding kappa-casein proteins are SEQ ID Nos. 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 and 148. The kappa-casein protein may be a kappa-casein protein from any mammalian species, for example a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, son, tiger, echidna or woolly mammoth kappa-casein protein. Additional sequences of different kappa-casein proteins and nucleic acids encoding different kappa-casein proteins are known in the art.

The kappa-casein protein may also be a protein that is at least 50% (e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type kappa-casein protein (e.g., SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, or 23). A nucleic acid encoding a kappa-casein protein may encode a protein that is at least 50% (e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type kappa-casein protein (e.g., SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, or 23).

alpha-S1-casein

Non-limiting examples of alpha-S1-casein proteins are SEQ ID Nos. 48, 50, 52, 54, 56, 57, 59, 61, 63, 64, 66, 68, 70, 72, 74 and 76. Non-limiting examples of nucleic acid sequences encoding alpha-S1-casein protein are SEQ ID Nos. 49, 51, 53, 55, 58, 60, 62, 65, 67, 69, 71, 73, 75, 77 and 147. The α -S1-casein protein may be an α -S1-casein protein from any mammalian species, for example, a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain goat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or mammoth α -S1-casein protein. Additional sequences of different alpha-S1-casein proteins and nucleic acids encoding different alpha-S1-casein proteins are known in the art.

The α -S1-casein protein may also be at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to the wild-type α -S1-casein protein (e.g., SEQ ID Nos: 48, 50, 52, 54, 56, 57, 59, 61, 63, 64, 66, 68, 70, 72, 74, or 76). A nucleic acid encoding an alpha-S1-casein protein may encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type alpha-S1-casein protein (e.g., SEQ ID Nos: 48, 50, 52, 54, 56, 57, 59, 61, 63, 64, 66, 68, 70, 72, 74, or 76).

alpha-S2-casein

Non-limiting examples of alpha-S2-casein proteins are SEQ ID Nos. 78, 80, 82, 84, 86, 88 and 90. Non-limiting examples of nucleic acid sequences encoding alpha-S2-casein proteins are SEQ ID Nos 79, 81, 83, 85, 87, 89, 91, 145 and 146. The α -S2-casein protein may be an α -S2-casein protein from any mammalian species, for example, a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain goat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or mammoth α -S2-casein protein. Additional sequences of different alpha-S2-casein proteins and nucleic acids encoding different alpha-S2-casein proteins are known in the art.

The α -S2-casein protein may also be at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type α -S2-casein protein (e.g., SEQ ID Nos: 78, 80, 82, 84, 86, 88, or 90). The nucleic acid encoding an alpha-S2-casein protein may encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type alpha-S2-casein protein (e.g., SEQ ID Nos: 78, 80, 82, 84, 86, 88, or 90).

Micelles comprising casein protein

In bovine milk, casein or casein micelles typically comprise 2.5% of the total mixture in suspension. If there is not enough casein, micelles will not form which are important for optimal performance of milk. Too much protein is not properly dissolved, resulting in an undesirable mixture. Casein micelles may include water and salts-mainly calcium and phosphorus. The casein micelles were easily separated and removed by centrifugation. It is easily separated from whey by precipitating casein with acid to lower the pH to about 4.6.

In some embodiments, the micelle may comprise a beta-casein protein (e.g., any beta-casein protein described herein) and a kappa-casein protein (e.g., any kappa-casein protein described herein). In some examples, the ratio of beta-casein protein to kappa-casein protein in the micelle is about 2.0:1 to about 5.5:1, 2.0:1 to about 5.0:1, 2.0:1 to about 4.5:1, about 2.0:1 to about 4.0:1, about 2.0:1 to about 3.5:1, about 2.0:1 to about 3.0:1, about 2.0:1 to about 2.5:1, about 2.5:1 to about 5.0:1, about 2.5:1 to about 4.5:1, about 2.5:1 to about 4.0:1, about 2.5:1 to about 3.5:1, about 2.5:1 to about 3.0:1, 3.0:1 to about 5.0:1, about 3.0:1 to about 4.5:1, about 3.0:1 to about 3.5:1, about 3.5:1 to about 3.0:1, about 3.5:1 to about 5:1, about 3.0:1, about 3.5:1, about 3.0:1 to about 5:1, about 3.0:1, about 5:1, about 5.0:1, about 5:1, about 5.0:1, about 5:1, about 4.0:1, about 5:1, about 5.0:1, about 5:1, about 5.0:1, about 5:1, about 5.0:1, about.

In some examples, the micelles have the following diameters (or micelle populations having the following average diameters): 20 nm to about 350 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, about 140 nm, about 120 nm, about 100 nm, about 80 nm, about 60 nm, or about 40 nm; about 40 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, about 140 nm, about 120 nm, about 100 nm, about 80 nm, or about 60 nm; about 60 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, about 140 nm, about 120 nm, or about 100 nm; about 80 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, about 140 nm, about 120 nm, or about 100 nm; about 100 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, about 140 nm, or about 120 nm; about 120 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, about 160 nm, or about 140 nm; about 140 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, about 180 nm, or about 160 nm; about 160 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, about 200 nm, or about 180 nm; about 180 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, about 220 nm, or about 200 nm; about 200 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, about 240 nm, or about 220 nm; about 220 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, about 260 nm, or about 240 nm; about 240 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, about 280 nm, or about 260 nm; about 260 nm to about 350 nm, about 340 nm, about 320 nm, about 300 nm, or about 280 nm; about 280 nm to about 350 nm, about 340 nm, about 320 nm, or about 300 nm; about 300 nm to about 350 nm or about 325 nm; or from about 325 nm to about 350 nm.

Whey protein

Whey is commonly referred to as a by-product of cheese and is also believed to be a cause of milk allergy. A typical whey composition comprises a mixture of beta-lactoglobulin, alpha-lactalbumin, serum albumin, immunoglobulins, lactoferrin, and transferrin. Whey proteins contain no phosphorus and remain in solution at low pH, whereas casein does not. In one embodiment, a selected combination of lactalbumin, comprising β -lactoglobulin and α -lactalbumin, is used as the major component or at least part of the milk protein component or composition. Non-limiting examples of different whey proteins are provided below.

Alpha-lactalbumin

Non-limiting examples of alpha-lactalbumin proteins are SEQ ID Nos 92, 94, 96 and 98. Non-limiting examples of nucleic acid sequences encoding alpha-lactalbumin proteins are SEQ ID NOs 93, 95, 97, 99 and 157. The alpha-lactalbumin protein may be an alpha-lactalbumin protein from any mammalian species, for example a bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat of north america (mountain goat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, possum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna or mammoth alpha-lactalbumin proteins. Additional sequences of different alpha-lactalbumin proteins and nucleic acids encoding different alpha-lactalbumin proteins are known in the art.

The alpha-lactalbumin protein may also be at least 80% (e.g. at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 99.5%) identical to the wild-type alpha-lactalbumin protein (e.g. SEQ ID Nos: 92, 94, 96 or 98). The nucleic acid encoding an alpha-lactalbumin protein may encode a protein that is at least 80% (e.g. at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 99.5%) identical to the wild-type alpha-lactalbumin protein (e.g. SEQ ID Nos: 92, 94, 96 or 98).

Beta-lactoglobulin

Non-limiting examples of beta-lactoglobulin proteins are SEQ ID Nos 100, 102, 104 and 106. Non-limiting examples of nucleic acid sequences encoding beta-lactoglobulin proteins are SEQ ID NOs 101, 103, 105, 107 and 143. The β -lactoglobulin protein may be a β -lactoglobulin protein from any mammalian species, for example, a bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american goat (mountain goat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth β -lactoglobulin protein. Additional sequences of different beta-lactoglobulin proteins and nucleic acids encoding different beta-lactoglobulin proteins are known in the art.

The beta-lactoglobulin protein may also be a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type beta-lactoglobulin protein (e.g., SEQ ID Nos: 100, 102, 104, or 106). The nucleic acid encoding a beta-lactoglobulin protein may encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type beta-lactoglobulin protein (e.g., SEQ ID Nos: 100, 102, 104, or 106).

Lactoferrin

Non-limiting examples of lactoferricin proteins are SEQ ID Nos. 108, 110, 112 and 114. Non-limiting examples of nucleic acid sequences encoding lactoferrin proteins are SEQ ID NOS 109, 111, 113 and 115. The lactoferrin protein can be lactoferrin from any mammalian species, for example, cow, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, marsupial (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, gill, or woolly crab. Additional sequences of different lactoferrins and nucleic acids encoding different lactoferrins are known in the art.

The lactoferricin protein may also be a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type lactoferricin protein (e.g., SEQ ID Nos: 108, 110, 112, or 114). The nucleic acid encoding a lactoferricin protein may encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type lactoferricin protein (e.g., SEQ ID Nos: 108, 110, 112, or 114).

Transferrin

Non-limiting examples of transferrin proteins are SEQ ID Nos. 116 and 118. Non-limiting examples of nucleic acid sequences encoding transferrin protein are SEQ ID NO 117 and 119. The transferrin protein can be a transferrin protein from any mammalian species, such as a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, marsupand (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, gill, or woolly down. Additional sequences of different transferrin proteins and nucleic acids encoding different transferrin proteins are known in the art.

The transferrin protein can also be a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type transferrin protein (e.g., SEQ ID Nos. 116 or 118). A nucleic acid encoding a transferrin protein can encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type transferrin protein (e.g., SEQ ID Nos: 116 or 118).

Serum albumin

Non-limiting examples of serum albumin proteins are SEQ ID Nos 120, 122, 124 and 126. Non-limiting examples of nucleic acid sequences encoding serum albumin proteins are SEQ ID NOs 121, 123, 125 and 127. The serum albumin protein may be a serum albumin protein from any mammalian species, such as bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, marsupillary (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, gill, or woolly down. Additional sequences of different serum albumin proteins and nucleic acids encoding different serum albumin proteins are known in the art.

The serum albumin protein can also be at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type serum albumin protein (e.g., SEQ ID Nos: 20, 122, 124, or 126). The nucleic acid encoding a serum albumin protein can encode a protein that is at least 80% (e.g., at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%) identical to a wild-type serum albumin protein (e.g., SEQ ID Nos: 20, 122, 124, or 126).

Lipids in mammalian produced milk

Milk fat contains about 400 different fatty acids, making it the most complex of all natural fats. Milk fatty acids are derived almost equally from two sources, feed and microbial activity in the rumen of cows, while the lipids in cow's milk are mainly present in the globules as oil-in-water emulsions. Fat is present in all natural dairy products and is essential for organoleptic characteristics such as flavour, mouthfeel and consistency. In addition, fats provide nutritional and health benefits.

Milk fat consists mainly of triglycerides, about 98%, while the other milk lipids are diacylglycerols (about 2% of the lipid fraction), cholesterol (less than 0.5%), phospholipids (about 1%) and Free Fatty Acids (FFA) (about 0.1%) Jensen RG, Newburg DS. Bovine milk lipids, Handbook of milk composition, Jensen RG London: Academic Press; 1995.543-75. furthermore, there are also traces of ether lipids, hydrocarbons, fat-soluble vitamins, flavors and feed-introduced compounds (Lindmark manson H., Food & Nutrition Research 2008. DOI: 10.3402/fnr. v52i0.1821).

Milk fatty triglycerides are synthesized from over 400 different fatty acids, making milk fat the most complex of all natural fats. Almost all fatty acids are present in trace amounts in milk, while only about 15 acids are present at levels of 1% or higher. Many factors are associated with changes in the amount of milk lipids and fatty acid composition. They may be of animal origin, i.e. associated with genetics (breed and selection), lactation, mastitis and rumen fermentation, or they may be feed-related factors, i.e. associated with fiber and energy intake, dietary fat and seasonal and regional effects. The fatty acids in milk fat are arranged in triglycerides according to a pattern that seems to be commonly used in ruminants. The percentage of unsaturated fatty acids (e.g., oleic acid and linolenic acid) in goats was not different from the average found for bovine milk. The main difference between the milk fat of goats and cows is the percentage distribution between specific short chain fatty acids. Goats have significantly higher capric, caprylic and caproic acids. The high amount of these specific fatty acids is responsible for the characteristic flavor and odor associated with goat milk. John C, Bruhn, FST, UC Davis, Davis, CA 95616-. Doi: 10.3402/fnr. v52i0.1821.

Milk fatty acids are derived almost equally from two sources, feed and microbial activity in the rumen of dairy cows. The fatty acid synthesis system in the mammary glands of cows produces fatty acids of even number of carbons in length between 4 and 16 and constitutes about 60% and 45% of the fatty acids on a molar and weight basis. This de novo synthesis in the mammary gland is 4: 0-14: 0 acid and approximately half of 16:0 is from acetate and beta-hydroxybutyrate. Acetic acid and butyric acid are produced in the rumen by fermentation of feed components. During absorption through the rumen epithelium, butyric acid is converted to beta-hydroxybutyrate.

Medium and long chain fatty acids, but predominantly 18:0, can be desaturated in the mammary gland to form the corresponding mono-saturated acids.

Fatty acids are not randomly esterified at the three positions of the triacylglycerol molecule(MacGibbon AHK, Taylor MW. Composition and structure of bone milk fine advanced design and design, Fox PFMcSweeney PLHNew York, Springer; 2006.1-42.). The short-chain acids butyric acid (4:0) and hexose (6:0) are almost completely esterified at sn-3. Medium chain fatty acids (8: 0-14: 0) and 16:0 are preferentially esterified at positions sn-1 and sn-2. Stearic acid (18:0) is selectively placed in position sn-1, while oleic acid (18:1) shows preference for positions sn-1 and sn-3 (Lindmark 2008).

There are milk replacement products that have a fat component formulated to the selected fatty acid characteristics, however, such triglycerides are not transesterified to long chain monounsaturated fatty acid triglycerides, such as found in vegetable oils. Us patent application No. 20140147548 discloses milk replacers for young animals by the addition of medium chain triglycerides, in particular caproic, caprylic, capric and lauric fatty acids or combinations thereof.

Lipids in the present compositions

The lipid in any composition or for use in any of the methods described herein can comprise: one or more fats, one or more oils, one or more mono-, di-and/or triglycerides, one or more free fatty acids and one or more phospholipids. Exemplary oils, monoglycerides, diglycerides, free fatty acids, and phospholipids are described below. Additional examples of fats, oils, monoglycerides, diglycerides, triglycerides, free fatty acids, and phospholipids are known in the art.

Oil

The oil used in the present compositions or methods may include, for example, oils of vegetable origin. Non-limiting examples of vegetable-based oils include sunflower oil, coconut oil, peanut oil, corn oil, cottonseed oil, olive oil, palm oil, rapeseed oil, safflower oil, sesame oil, soybean oil, almond oil, beech nut oil, brazil nut oil, cashew nut oil, hazelnut oil, macadamia nut oil, Mongolian nut oil, pecan oil, pine nut oil, pistachio nut oil, walnut oil, and avocado oil.

Mono-and diglycerides

The mono-and diglycerides which can be used in the present invention may be those of vegetable origin. For example, the monoglycerides and diglycerides may be derived from sunflower, coconut, peanut, cottonseed, olive, palm, rapeseed, safflower, sesame seed, soybean, almond, beech nut, brazil nut, cashew, hazelnut, macadamia nut, mongolian nut, pecan, pine nut, pistachio, walnut oil, and avocado. The monoglycerides and diglycerides may include the acyl chains of any of the free fatty acids listed herein. Additional examples of monoglycerides and diglycerides are known in the art.

Free fatty acids

The compositions described herein may include and the methods described herein may include the use of one or more free fatty acids. Non-limiting examples of free fatty acids include butyric acid, caproic acid, caprylic acid, and capric acid. Additional examples of fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, myristoleic acid, palmitoleic acid (pamitoleic acid), sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, oleic acid, linolenic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, omega-3 fatty acids, and omega-6 fatty acids. In some examples, the free fatty acids are saturated. In some examples, the free fatty acid is unsaturated. In some embodiments, the free fatty acids are not derived from or produced by a mammal. Additional examples of free fatty acids are known in the art.

Phospholipids

The compositions described herein and the methods described herein may include the use of one or more phospholipids. Non-limiting examples of phospholipids include lecithin phospholipids (e.g., soybean lecithin phospholipids, sunflower lecithin phospholipids, cotton lecithin phospholipids, rapeseed lecithin phospholipids, rice bran lecithin phospholipids, and corn lecithin phospholipids). In some embodiments, the phospholipid is not derived from or produced by a mammal. Additional aspects of phospholipids are known in the art.

Flavour compounds

Any of the compositions or methods described herein may include or include the use of one or more different flavor compounds. Non-limiting examples of flavor compounds include delta-decalactone, ethyl butyrate, 2-furylmethyl ketone, 2, 3-pentanedione, gamma undecalactone, and delta undecalactone. Further examples of flavour compounds include artificial flavours such as chocolate, coffee, strawberry, almond, hazelnut, vanilla, green tea, irish cream and coconut flavour. Additional examples of flavor compounds are known in the art.

Ash content

Any of the compositions or methods described herein may include or include the use of ash. The ash may, for example, comprise one or more (two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty) of the following: calcium, phosphorus, potassium, sodium, citrate, chloride, phosphate, magnesium, iron, molybdenum, manganese, copper, thiamine (vitamin B1), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), vitamin B6 (pyridoxine), vitamin B12 (cobalamin), vitamin C, folate, vitamin a, vitamin D, vitamin E, and vitamins. In some examples, the ash includes CaCl ₂、KH₂PO₄And sodium citrate (e.g., two or three). The ash may be provided as a powder or a solution. Additional components and aspects of ash are known in the art. In some embodiments, the ash is not derived from or produced by a mammal.

Colour balancing agent

Various color balancing agents are known in the art. For example, the color balancing agent can be a compound obtained from a plant (e.g., a monocot or a dicot). In some examples, the color balancing agent is a synthetic compound. In some examples, the color-balancing agent is not obtained from or produced by a mammal or mammalian cell. Non-limiting examples of color balancing agents include beta-carotene and annatto.

Sweetening agent

The sweetener may be a sugar (e.g., a monosaccharide, disaccharide, or polysaccharide) or an artificial sweetener. Non-limiting examples of sweeteners as sugars include glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, and tagatose. Additional examples of sugars that can be used as sweeteners in any of the compositions or methods described herein are known in the art.

Non-limiting examples of sweeteners as artificial sweeteners include stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osthole, betadine, sorbitol, thaumatin, xylitol, potassium acetaminosulfonate, advatame, alitame, aspartame-acesulfame, sodium cyclamate, dulcin, neohesperidin dihydrochalcone, neotame, and P-4000. Additional artificial sweeteners useful as sweeteners in any of the compositions or methods described herein are known in the art.

Composition comprising a metal oxide and a metal oxide

Provided herein are compositions comprising: about 0.3 g/L to about 1.1 g/L (e.g., about 0.3 g/L to about 1.0 g/L, about 0.3 g/L to about 0.9 g/L, about 0.3 g/L to about 0.8 g/L, about 0.3 g/L to about 0.7 g/L, about 0.3 g/L to about 0.6 g/L, about 0.3 g/L to about 0.5 g/L, about 0.3 g/L to about 0.4 g/L, about 0.4 g/L to about 1.1 g/L, about 0.4 g/L to about 1.0 g/L, about 0.4 g/L to about 0.9 g/L, about 0.4 g/L to about 0.8 g/L, about 0.4 g/L to about 0.7 g/L, about 0.4 g/L to about 0.6 g/L, about 0.4 g/L to about 0.5 g/L, about 0.4 g/L, About 0.5 g/L to about 1.1 g/L, about 0.5 g/L to about 1.0 g/L, about 0.5 g/L to about 0.9 g/L, about 0.5 g/L to about 0.8 g/L, about 0.5 g/L to about 0.7 g/L, about 0.5 g/L to about 0.6 g/L, about 0.6 g/L to about 1.1 g/L, about 0.6 g/L to about 1.0 g/L, about 0.6 g/L to about 0.9 g/L, about 0.6 g/L to about 0.8 g/L, about 0.6 g/L to about 0.7 g/L, about 0.7 g/L to about 1.1 g/L, about 0.7 g/L to about 1.0 g/L, about 0.7 g/L to about 0.9 g/L, about 0.8 g/L, about 0.7 g/L to about 0.8 g/L, About 0.8 g/L to about 1.1 g/L, about 0.8 g/L to about 1.0 g/L, about 0.8 g/L to about 0.9 g/L, about 0.9 g/L to about 1.1 g/L, about 0.9 g/L to about 1.0 g/L, about 1.0 g/L to about 1.1 g/L, or about 0.27 wt% to about 0.75 wt%) of a kappa-casein protein (e.g., any of the kappa-casein proteins described herein); about 1.25 g/L to about 4.9 g/L (e.g., about 1.25 g/L to about 4.6 g/L, about 1.25 g/L to about 4.4 g/L, about 1.25 g/L to about 4.2 g/L, about 1.25 g/L to about 4.0 g/L, about 1.25 g/L to about 3.8 g/L, about 1.25 g/L to about 3.6 g/L, about 1.25 g/L to about 3.4 g/L, about 1.25 g/L to about 3.2 g/L, about 1.25 g/L to about 3.0 g/L, about 1.25 g/L to about 2.8 g/L, about 1.25 g/L to about 2.6 g/L, about 1.25 g/L to about 2.4 g/L, about 1.25 g/L to about 2.2 g/L, about 1.2 g/L to about 2.0 g/L, about 1.25 g/L to about 2.0 g/L, About 1.25 g/L to about 1.8 g/L, about 1.25 g/L to about 1.6 g/L, about 1.25 g/L to about 1.4 g/L, about 1.4 g/L to about 4.9 g/L, about 1.4 g/L to about 4.6 g/L, about 1.4 g/L to about 4.4 g/L, about 1.4 g/L to about 4.2 g/L, about 1.4 g/L to about 4.0 g/L, about 1.4 g/L to about 3.8 g/L, about 1.4 g/L to about 3.6 g/L, about 1.4 g/L to about 3.4 g/L, about 1.4 g/L to about 3.2 g/L, about 1.4 g/L to about 3.0 g/L, about 1.4 g/L to about 2.8 g/L, about 1.4 g/L to about 2 g/L, about 1.4 g/L to about 3.0 g/L, About 1.4 g/L to about 2.4 g/L, about 1.4 g/L to about 2.2 g/L, about 1.4 g/L to about 2.0 g/L, about 1.4 g/L to about 1.8 g/L, about 1.4 g/L to about 1.6 g/L, about 1.6 g/L to about 4.9 g/L, about 1.6 g/L to about 4.6 g/L, about 1.6 g/L to about 4.4 g/L, about 1.6 g/L to about 4.2 g/L, about 1.6 g/L to about 4.0 g/L, about 1.6 g/L to about 3.8 g/L, about 1.6 g/L to about 3.6 g/L, about 1.6 g/L to about 3.4 g/L, about 1.6 g/L to about 3.2 g/L, about 1.6 g/L to about 3.6 g/L, About 1.6 g/L to about 2.8 g/L, 1.6 g/L to about 2.6 g/L, about 1.6 g/L to about 2.4 g/L, about 1.6 g/L to about 2.2 g/L, about 1.6 g/L to about 2.0 g/L, about 1.6 g/L to about 1.8 g/L, about 1.8 g/L to about 4.9 g/L, about 1.8 g/L to about 4.6 g/L, about 1.8 g/L to about 4.4 g/L, about 1.8 g/L to about 4.2 g/L, about 1.8 g/L to about 4.0 g/L, about 1.8 g/L to about 3.8 g/L, about 1.8 g/L to about 3.6 g/L, about 1.8 g/L to about 3.4 g/L, about 1.8 g/L to about 3.8 g/L, about 1.8 g/L to about 2 g/L, About 1.8 g/L to about 3.0 g/L, about 1.8 g/L to about 2.8 g/L, 1.8 g/L to about 2.6 g/L, about 1.8 g/L to about 2.4 g/L, about 1.8 g/L to about 2.2 g/L, about 1.8 g/L to about 2.0 g/L, about 2.0 g/L to about 4.9 g/L, about 2.0 g/L to about 4.6 g/L, about 2.0 g/L to about 4.4 g/L, about 2.0 g/L to about 4.2 g/L, about 2.0 g/L to about 4.0 g/L, about 2.0 g/L to about 3.8 g/L, about 2.0 g/L to about 3.6 g/L, about 2.0 g/L to about 3.4 g/L, about 2.0 g/L to about 2.0 g/L, about 2.0 g/L to about 2.4 g/L, about 2 g/L to about 2.2 g/L, About 2.0 g/L to about 3.0 g/L, about 2.0 g/L to about 2.8 g/L, 2.0 g/L to about 2.6 g/L, about 2.0 g/L to about 2.4 g/L, about 2.0 g/L to about 2.2 g/L, about 2.2 g/L to about 4.9 g/L, about 2.2 g/L to about 4.6 g/L, about 2.2 g/L to about 4.4 g/L, about 2.2 g/L to about 4.2 g/L, about 2.2 g/L to about 4.0 g/L, about 2.2 g/L to about 3.8 g/L, about 2.2 g/L to about 3.6 g/L, about 2.2 g/L to about 3.4 g/L, about 2.2 g/L to about 3.2 g/L, about 2 g/L to about 2.2 g/L to about 3.2 g/L, About 2.2 g/L to about 2.8 g/L, 2.2 g/L to about 2.6 g/L, about 2.2 g/L to about 2.4 g/L, about 2.4 g/L to about 4.9 g/L, about 2.4 g/L to about 4.6 g/L, about 2.4 g/L to about 4.4 g/L, about 2.4 g/L to about 4.2 g/L, about 2.4 g/L to about 4.0 g/L, about 2.4 g/L to about 3.8 g/L, about 2.4 g/L to about 3.6 g/L, about 2.4 g/L to about 3.4 g/L, about 2.4 g/L to about 3.2 g/L, about 2.4 g/L to about 3.0 g/L, about 2.4 g/L to about 2.8 g/L, about 2.4 g/L to about 2.6 g/L, About 2.6 g/L to about 4.9 g/L, about 2.6 g/L to about 4.6 g/L, about 2.6 g/L to about 4.4 g/L, about 2.6 g/L to about 4.2 g/L, about 2.6 g/L to about 4.0 g/L, about 2.6 g/L to about 3.8 g/L, about 2.6 g/L to about 3.6 g/L, about 2.6 g/L to about 3.4 g/L, about 2.6 g/L to about 3.2 g/L, about 2.6 g/L to about 3.0 g/L, about 2.6 g/L to about 2.8 g/L, about 2.8 g/L to about 4.9 g/L, about 2.8 g/L to about 4.6 g/L, about 2.8 g/L to about 4.8 g/L, about 2.8 g/L to about 4.6 g/L, about 2.8 g/L to about 2.8 g/L, About 2.8 g/L to about 4.0 g/L, about 2.8 g/L to about 3.8 g/L, about 2.8 g/L to about 3.6 g/L, about 2.8 g/L to about 3.4 g/L, about 2.8 g/L to about 3.2 g/L, about 2.8 g/L to about 3.0 g/L, about 3.0 g/L to about 4.9 g/L, about 3.0 g/L to about 4.6 g/L, about 3.0 g/L to about 4.4 g/L, about 3.0 g/L to about 4.2 g/L, about 3.0 g/L to about 4.0 g/L, about 3.0 g/L to about 3.8 g/L, about 3.0 g/L to about 3.6 g/L, about 3.0 g/L to about 3.4 g/L, about 3.0 g/L to about 3.0 g/L, about 3.3.0 g/L to about 3.8 g/L, About 3.2 g/L to about 4.9 g/L, about 3.2 g/L to about 4.6 g/L, about 3.2 g/L to about 4.4 g/L, about 3.2 g/L to about 4.2 g/L, about 3.2 g/L to about 4.0 g/L, about 3.2 g/L to about 3.8 g/L, about 3.2 g/L to about 3.6 g/L, about 3.2 g/L to about 3.4 g/L, about 3.4 g/L to about 4.9 g/L, about 3.4 g/L to about 4.6 g/L, about 3.4 g/L to about 4.4 g/L, about 3.4 g/L to about 4.2 g/L, about 3.4 g/L to about 4.0 g/L, about 3.4 g/L to about 3.8 g/L, about 3.4 g/L to about 3.6 g/L, About 3.6 g/L to about 4.9 g/L, about 3.6 g/L to about 4.6 g/L, about 3.6 g/L to about 4.4 g/L, about 3.6 g/L to about 4.2 g/L, about 3.6 g/L to about 4.0 g/L, about 3.6 g/L to about 3.8 g/L, about 3.8 g/L to about 4.9 g/L, about 3.8 g/L to about 4.6 g/L, about 3.8 g/L to about 4.4 g/L, about 3.8 g/L to about 4.2 g/L, about 3.8 g/L to about 4.0 g/L, about 4.0 g/L to about 4.9 g/L, about 4.0 g/L to about 4.6 g/L, about 4.0 g/L to about 4.4 g/L, about 4.0 g/L to about 4.0 g/L, about 4.0 g/L to about 4.2 g/L, About 4.2 g/L to about 4.9 g/L, about 4.2 g/L to about 4.6 g/L, about 4.2 g/L to about 4.4 g/L, about 4.4 g/L to about 4.9 g/L, about 4.4 g/L to about 4.6 g/L, about 4,6 g/L to about 4.9 g/L, or about 1.23 wt% to about 3.27 wt%) beta-casein protein (e.g., any of the beta-casein proteins described herein); the final total concentration is about 0 wt% to about 45 wt% (e.g., 0 wt%, about 0 wt% to about 4.5 wt%, about 0.5 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 5 wt%, about 4 wt%, about 3 wt%, about 2 wt%, or about 1 wt%, about 1.0 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 5 wt%, about 4 wt%, about 3 wt%, or about 2 wt%, about 2 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 25 wt%, about 20 wt%, or about 2 wt%, About 10 wt%, about 8 wt%, about 6 wt%, about 5 wt%, about 4 wt%, or about 3 wt%; about 3 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 5 wt%, or about 4 wt%; about 4 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, about 8 wt%, about 6 wt%, or about 5 wt%; about 5 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, about 8 wt%, or about 6 wt%; about 6 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, about 10 wt%, or about 8 wt%; about 8 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, about 15 wt%, or about 10 wt%; about 10 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, about 20 wt%, or about 15 wt%; about 15 wt% to about 40 wt%, about 35 wt%, about 30 wt%, about 25 wt%, or about 20 wt%; about 20 wt% to about 40 wt%, about 35 wt%, about 30 wt%, or about 25 wt%; about 25 wt% to about 40 wt%, about 35 wt%, or about 30 wt%; about 30 wt% to about 40 wt%, or about 35 wt%; or about 35% to about 40% by weight) of one or more lipids (e.g., any one or more lipids described herein); the final total concentration is about 0.01 wt% to about 6 wt% (e.g., about 0.1 wt% to about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, about 2.0 wt%, about 1.5 wt%, about 1.0 wt%, or about 0.5 wt%, (about 0.5 wt% to about 6 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, about 2.0 wt%, about 1.5 wt%, or about 1.0 wt%, (about 1.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.0 wt%, about 3.5 wt%, about 1.5 wt%, about 1.0 wt%, about 5.5 wt%, about 2.0 wt%, about 5.5 wt%, about 5.0 wt%, about 5 wt%, or about 5.5 wt%, About 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, or about 2.0 wt%; about 2.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, or about 2.5 wt%; about 2.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, or about 3.0 wt%; about 3.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, or about 3.5 wt%; about 3.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, or about 4.0 wt%; about 4.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%; about 4.5 wt% to about 6.0 wt%, about 5.5 wt%, or about 5.0 wt%; about 5.0 wt% to about 6.0 wt% or about 5.5 wt%; or from about 5.5% to about 6.0% by weight) of one or more flavor compounds (e.g., any one or more flavor compounds described herein); the final total concentration is about 0.1 wt% to about 6 wt% (e.g., about 0.1 wt% to about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, about 2.0 wt%, about 1.5 wt%, about 1.0 wt%, or about 0.5 wt%; about 0.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, about 2.0 wt%, about 1.5 wt%, or about 1.0 wt%, about 1.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.0 wt%, about 3.5 wt%, about 1.0 wt%, about 1.5 wt%, about 2.0 wt%, about 5.5 wt%, about 5.0 wt%, about 5 wt%, or about 5.0 wt%, about 5 wt%, about 2.0 wt%, about 5 wt%, or about 5.5 wt%, About 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, about 2.5 wt%, or about 2.0 wt%; about 2.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 3.0 wt%, or about 2.5 wt%; about 2.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, or about 3.0 wt%; about 3.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, or about 3.5 wt%; about 3.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, or about 4.0 wt%; about 4.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, or about 4.5 wt%; about 4.5 wt% to about 6.0 wt%, about 5.5 wt%, or about 5.0 wt%; about 5.0 wt% to about 6.0 wt%, or about 5.5 wt%; or about 5.5% to about 6.0% by weight) of one or more sweeteners (e.g., any one or more of the sweeteners described herein); and a final total concentration of about 0.15 wt% to about 1.5 wt% (e.g., about 0.15 wt% to about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.6 wt%, about 0.5 wt%, about 0.4 wt%, about 0.3 wt%, or about 0.2 wt%, about 0.2 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.6 wt%, about 0.5 wt%, about 0.4 wt%, or about 0.3 wt%, about 0.3 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.0 wt%, about 1.4 wt%, about 1.1.6 wt%, about 0.5 wt%, about 0.4 wt%, about 0.1.1 wt%, about 0.1.4 wt%, about 0.3 wt%, about 0.5 wt%, about 0.9 wt%, about 0.4 wt%, about 0.1 wt%, about 0.1.1 wt%, about 0.4 wt%, about 0.1 wt%, or about 0.3 wt%, About 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.6 wt%, or about 0.5 wt%; about 0.5 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, or about 0.6 wt%; about 0.6 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, about 0.9 wt%, or about 0.8 wt%; about 0.8 wt% to about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, about 1.0 wt%, or about 0.9 wt%; about 0.9 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, about 1.1 wt%, or about 1.0 wt%; about 1.0 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, about 1.2 wt%, or about 1.1 wt%; about 1.1 wt% to about 1.5 wt%, about 1.4 wt%, about 1.3 wt%, or about 1.2 wt%; about 1.2 wt% to about 1.5 wt%, about 1.4 wt%, or about 1.3 wt%; about 1.3 wt% to about 1.5 wt% or about 1.4 wt%; or from about 1.4 wt.% to about 1.5 wt.%) ash, wherein the composition does not include animal-derived components.

Also provided are compositions comprising: about 0.3g/L to about 1.1g/L (e.g., any subrange from about 0.3g/L to about 1.1g/L described in the preceding paragraph) kappa-casein protein (e.g., any kappa-casein protein described herein); about 1.25 g/L to about 4.9 g/L (e.g., any subrange from about 1.25 g/L to about 4.9 g/L described in the preceding paragraph) beta-casein protein (e.g., any beta-casein protein described herein); a final total concentration of one or more lipids (e.g., any one or more lipids described herein) of about 0 wt% to about 45 wt% (e.g., all subranges from about 0 wt% to about 45 wt% described in the above paragraph); a final total concentration of one or more flavor compounds (e.g., any one or more flavor compounds described herein) of about 0.01 wt% to about 6 wt% (e.g., all subranges from about 0.01 wt% to about 6 wt% described in the above paragraph); (ii) one or more sweeteners (e.g., any one or more of the sweeteners described herein) at a final total concentration of about 0.1% to about 6% by weight (e.g., all subranges from about 0.1% to about 6% by weight described in the above paragraph); and an ash (e.g., any of the exemplary ashes described herein) at a final total concentration of about 0.15 wt.% to about 1.5 wt.% (e.g., all subranges of about 0.15 wt.% to about 1.5 wt.% described in the above paragraph), wherein: the composition is as follows: excluding at least one component found in milk produced by a mammal; comprising at least one component not present in mammalian-produced milk; and/or comprises at least one component at a higher or lower concentration than the concentration of at least one component in milk produced by the mammal. In some examples of these compositions, the composition comprises a higher concentration of at least one component compared to the concentration of one or more components in milk produced by the mammal, the component selected from the group consisting of: calcium, phosphate, B complex vitamin, vitamin A, vitamin D, vitamin E and vitamin K. In some embodiments of these compositions, the composition does not include at least one component found in milk produced by a mammal selected from the group consisting of: lactose, bacteria, mycobacteria, allergens, viruses, prions, yeast, growth hormones, leukocytes, antibiotics, heavy metals, immunoglobulins, lactoferrin, lactoperoxidase and lipase. In some examples of these compositions, the composition comprises at least one component not present in mammalian-produced milk selected from the group consisting of artificial sweeteners, plant-derived lipids, β -casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern, and κ -casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern.

Also provided are compositions comprising: about 0.3g/L to about 1.1g/L (e.g., any subrange from about 0.3g/L to about 1.1g/L described in this section) a kappa-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern (e.g., any kappa-casein protein described herein); about 1.25 g/L to about 4.9 g/L (e.g., any subrange from about 1.25 g/L to about 4.9 g/L described in this section) β -casein protein that is not glycosylated or has a non-mammalian glycosylation pattern (e.g., any β -casein protein described herein); a final total concentration of about 0 wt% to about 45 wt% (e.g., all subranges from about 0 wt% to about 45 wt% described in this section) of one or more lipids (e.g., any one or more lipids described herein); a final total concentration of one or more flavor compounds (e.g., any one or more flavor compounds described herein) of about 0.01 wt% to about 6 wt% (e.g., all subranges from about 0.01 wt% to about 6 wt% described in this section); one or more sweeteners (e.g., any one or more of the sweeteners described herein) at a final total concentration of about 0.1% to about 6% by weight (e.g., all subranges from about 0.1% to about 6% by weight described in this section); and ash (e.g., any ash described herein) at a final total concentration of about 0.15 wt% to about 1.5 wt% (e.g., all subranges from about 0.15 wt% to about 1.5 wt% described in this section).

Also provided are compositions comprising micelles comprising a kappa-casein protein (e.g., any of the kappa-casein proteins described herein) and a beta-casein protein (e.g., any of the beta-casein proteins described herein), wherein the micelles have a diameter of about 50nm to about 350nm (e.g., any subrange of the diameter of the micelles described herein), and the kappa-casein protein and the beta-casein protein are non-glycosylated or have a non-mammalian glycosylation pattern. In some embodiments, the composition comprises micelles at a final concentration of: about 2.0 wt.% to about 6 wt.% (e.g., about 2.0 wt.% to about 5.5 wt.%, about 5.0 wt.%, about 4.5 wt.%, about 4.0 wt.%, about 3.5 wt.%, about 3.0 wt.%, or about 2.5 wt.%, about 2.5 wt.% to about 6.0 wt.%, about 5.5 wt.%, about 5.0 wt.%, about 4.5 wt.%, about 4.0 wt.%, about 3.5 wt.%, or about 3.0 wt.%, about 3.0 wt.% to about 6.0 wt.%, about 5.5 wt.%, about 5.0 wt.%, about 4.5 wt.%, about 4.0 wt.%, or about 3.5 wt.%, about 3.5 wt.% to about 6.0 wt.%, about 5.5 wt.%, about 5.0 wt.%, about 4.5 wt.%, about 4.0 wt.%, or about 3.5 wt.%, about 6.0 wt.%, about 5.5 wt.%, about 5.0 wt.%, about 5.5 wt.%, about 4.5 wt.%, about 4.0 wt.%, or about 3.5 wt.% (about 3.5 wt.% to about 6.0 wt.%), about 5 wt.%, Or about 4.0 wt%; about 4.5 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, about 4.5 wt%, or about 4.0 wt%; about 4.0 wt% to about 6.0 wt%, about 5.5 wt%, about 5.0 wt%, or about 4.5 wt%; about 4.5 wt% to about 5.5 wt%, or about 5.0 wt%; about 5.0 wt% to about 6.0 wt% or 5.5 wt%; or from about 5.5 wt% to about 6.0 wt%). In some embodiments of these compositions, the ratio of β -casein protein to κ -casein protein in the micelles is from about 2.0:1 to about 5.5:1 (e.g., any subrange of the ratio from about 2.0:1 to about 5.5:1 described for the micelles herein). In some embodiments, these compositions further comprise: a final total concentration of about 0 wt% to about 45 wt% (e.g., all subranges from about 0 wt% to about 45 wt% described in this section) of one or more lipids (e.g., any one or more lipids described herein); a final total concentration of one or more flavor compounds (e.g., any one or more flavor compounds described herein) of about 0.01 wt% to about 6 wt% (e.g., all subranges from 0.01 wt% to about 6 wt% described in this section); one or more sweeteners (e.g., any one or more of the sweeteners described herein) at a final total concentration of about 0.1% to about 6% by weight (e.g., all subranges from about 0.1% to about 6% by weight described in this section); and ash (e.g., any ash described herein) at a final total concentration of about 0.15 wt% to about 1.5 wt% (e.g., all subranges from about 0.15 wt% to about 1.5 wt% described in this section).

In some embodiments of any of the compositions described herein, the one or more lipids are selected from the group consisting of: sunflower oil, coconut oil, tributyrin, mono-and diglycerides, free fatty acids and phospholipids. Some examples of any of the compositions described herein further include one or more of the following: the final concentration is about 1 wt% to about 28 wt% (e.g., about 1 wt% to about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 4 wt%, or about 2 wt%, about 2 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, about 10 wt%, about 8 wt%, about 6 wt%, or about 4 wt%, about 4 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, about 10 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, about 10 wt%, About 8 wt%, or about 6 wt%; about 6 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, about 10 wt%, or about 8 wt%; about 8 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, about 12 wt%, or about 10 wt%; about 10 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, about 14 wt%, or about 12 wt%; about 12 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, about 16 wt%, or about 14 wt%; about 14 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%, or about 16 wt%; about 16 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, about 20 wt%, about 18 wt%; about 18 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%, or about 20 wt%; about 20 wt% to about 28 wt%, about 26 wt%, about 24 wt%, about 22 wt%; about 22 wt% to about 28 wt%, about 26 wt%, about 24 wt%; about 24 wt% to about 28 wt% or about 26 wt%; or from about 28 wt.% to about 30 wt.%) sunflower oil; a final concentration of about 0.5 wt% to about 14 wt% (e.g., about 0.5 wt% to about 12 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 4 wt%, about 2 wt%, or about 1 wt%, about 1 wt% to about 14 wt%, about 12 wt%, about 10 wt%, about 8 wt%, about 6 wt%, about 4 wt%, or about 2 wt%, about 2 wt% to about 12 wt%, about 10 wt%, about 8 wt%, about 6 wt%, or about 4 wt%, about 4 wt% to about 14 wt%, about 12 wt%, about 10 wt%, about 8 wt%, or about 6 wt%, about 6 wt% to about 14 wt%, about 12 wt%, about 10 wt%, or about 8 wt%, about 8 wt% to about 14 wt%, about 12 wt%, or about 10 wt%, about 10 wt% to about 14 wt%, or about 12 wt%, or about 14 wt% to about 12 wt%, or about 14 wt%, or about 12 wt%; the final concentration is from about 0.05 wt% to about 1.0 wt% (e.g., about 0.05 wt% to about 0.9 wt%, about 0.8 wt%, about 0.7 wt%, about 0.6 wt%, about 0.5 wt%, about 0.4 wt%, about 0.3 wt%, or about 0.2 wt%, 0.1 wt% to about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.7 wt%, about 0.6 wt%, about 0.5 wt%, about 0.4 wt%, about 0.3 wt%, or about 0.2 wt%, about 0.2 wt% to about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.7 wt%, about 0.6 wt%, about 0.5 wt%, or about 0.4 wt%, about 0.4 wt% to about 1.0.0 wt%, about 0.0 wt%, about 0.0.0 wt%, about 0.9 wt%, about 0.8 wt%, about 0.7 wt%, about 0.6 wt%, about 0.5 wt%, about 0.7 wt%, or about 0.7 wt%, about 0.6 wt%, about 0.5 wt%, or about 0.2 wt%, about 0.7 wt%, or about 0.4 wt%, about 0.7 wt%, or about 0.7 wt%, about 0.0.0.7 wt%, or about 0.7 wt%, about 0.0.0.0.9 wt%, about 0.7 wt%, or about 0.7 wt%, about 0.6 wt%, about 0.0.0.0.0.0.9 wt%, about 0.9 wt%, about 0.7 wt%, about 0.9 wt%, about 0.0.7 wt%, about 0.7 wt%, about 0.0.0.0.0.7 wt%, about 0.0.0.9 wt%, about 0.7 wt%, about 0.0.6 wt%, about 0.7 wt%, about 0.0.9 wt%, about 0.9 wt%, about 0.0.7 wt%, about 0.5 wt%, about 0.9 wt%, about 0.6 wt%, about 0.9 wt%, about 0.7 wt%, about 0.9 wt%, about 0.5 wt%, about 0.7 wt%, about 0.0.0.9 wt%, about 0.0.0.7 wt%, about 0.0.0.0.7 wt%, about 0.9 wt%, about 0.7 wt%, about 0.9 wt%, about 0.8 wt%, about 0.7 wt%, about 0.5 wt%, about 0.8 wt%, about 0, Or about 0.6 wt%; about 0.6 wt% to about 1.0 wt%, about 0.9 wt%, about 0.8 wt%, or about 0.7 wt%; about 0.7 wt% to about 1.0 wt%, about 0.9 wt%, or about 0.8 wt%; about 0.8 wt% to about 1.0 wt% or about 0.9 wt%; or from about 0.9 wt% to about 1.0 wt%) of tributyrin;

A final concentration of about 0.08 wt% to about 1.2 wt% (e.g., 0.08 wt% to about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, about 0.4 wt%, or about 0.2 wt%, about 0.2 wt% to about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, or about 0.4 wt%, about 0.4 wt% to about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, or about 0.6 wt%, about 0.6 wt% to about 1.2 wt%, about 1.0 wt%, or about 0.8 wt%, about 0.8 wt% to about 1.2 wt%, or about 1.0 wt% of a monoglyceride and diglyceride (e.g., any one or more of the monoglycerides or diglycerides described herein), or about 1.0 wt%) and a diglyceride; and a final total concentration of free fatty acids of about 0.02 wt% to about 0.28 wt%; and a final total concentration of phospholipids (e.g., any one or more of the phospholipids described herein) in a range of about 0.02 wt% to about 0.3 wt% (e.g., about 0.02 wt% to about 0.25 wt%, about 0.20 wt%, about 0.15 wt%, or about 0.10 wt%, about 0.05 wt% to about 0.3 wt%, about 0.25 wt%, about 0.20 wt%, about 0.15 wt%, or about 0.10 wt%, about 0.10 wt% to about 0.30 wt%, about 0.25 wt%, about 0.20 wt%, or about 0.15 wt%, about 0.15 wt% to about 0.30 wt%, about 0.25 wt%, or about 0.20 wt%, about 0.20 wt% to about 0.30 wt%, or about 0.25 wt% to about 0.30 wt%).

In some embodiments of any of the compositions, the free fatty acids include at least one (e.g., two, three, or four) fatty acid selected from: butyric acid, caproic acid, caprylic acid and capric acid. In some embodiments of any of the compositions, the phospholipid is soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. In some examples of any of the compositions described herein, the flavor compounds comprise at least one flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone. In some embodiments of any of the compositions described herein, the one or more sweeteners is a sugar (e.g., glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, or tagatose). In some examples of any of the compositions described herein, the one or more sweeteners are artificial sweeteners (e.g., stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, ostwald, betadine, sorbitol, thaumatin, xylitol, potassium acetaminosulfonate, advatame, alitame, aspartame-acetyl sulfamate, sodium cyclamate, dulcin, neohesperidin dihydrochalcone, neotame, or P-4000).

In some examples of any of the compositions described herein, the ash comprises one or more (e.g., two, three, four, five, or six) of: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some embodiments of any of the compositions described herein, the ash comprises CaCl₂、KH₂PO₄And sodium citrate (e.g., two or three). Some embodiments of the compositions described herein include: a final concentration of CaCl of about 0.05 to about 0.2 g/L (e.g., about 0.05 to about 0.15 g/L, about 0.05 to about 0.10 g/L, about 0.10 to about 0.20 g/L, about 0.10 to about 0.15 g/L, or about 0.15 to about 0.2 g/L)₂(ii) a A final concentration of about 0.2 g/L to about 0.4 g/L (e.g., about 0.2 g/L to about 0.35 g/L, about 0.2 g/L to about 0.30 g/L, about 0.2 g/L to about 0.25 g/L, about 0.25 g/L to about 0.4 g/L, about 0.25 g/L to about 0.30 g/L, about 0.30 g/L to about 0.40 g/L, or about 0.30 g/L to about 0.35 g/L, or about 0.35 g/L to about 0.40 g/L) KH₂PO₄(ii) a And/or sodium citrate at a final concentration of about 0.1 g/L to about 0.3 g/L (e.g., 0.1 g/L to about 0.25 g/L, about 0.1 g/L to about 0.20 g/L, about 0.1 g/L to about 0.15 g/L, about 0.15 g/L to about 0.30 g/L, about 0.15 g/L to about 0.25 g/L, about 0.15 g/L to about 0.20 g/L, about 0.20 g/L to about 0.30 g/L, about 0.20 g/L to about 0.25 g/L, or about 0.25 g/L to about 0.30 g/L).

In any of the compositions described herein, the kappa-casein protein may be a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly echis kappa-casein protein. In any of the compositions described herein, the β -casein protein may be a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly.

In some examples of any of the compositions described herein, it may further comprise: a final concentration of about 0.4 wt% to about 2.5 wt% (e.g., about 0.4 wt% to about 2.0 wt%, about 1.5 wt%, or about 1.0 wt%, about 1.0 wt% to about 2.5 wt%, about 2.0 wt%, or about 1.5 wt%, about 1.5 wt% to about 2.5 wt% or 2.0 wt%, or about 2.0 wt% to about 2.5 wt%) of an alpha-lactalbumin protein (e.g., any alpha-lactalbumin protein described herein), and/or a final concentration of about 2.5 wt% to about 4.5 wt% of a beta-lactoglobulin protein (e.g., any beta-lactoglobulin protein described herein). In some embodiments of any of the compositions described herein, the alpha-lactalbumin protein may be a bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, son, tiger, echidna, or woolly mammoth alpha-lactalbumin protein. In some embodiments of any of the compositions described herein, the β -lactoglobulin protein can be a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american wildcat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, oran, mandrill (mandrill), pig, wolf, fox, son, tiger, echidna, or woolly mammoth β -lactoglobulin protein.

Some embodiments of any of the compositions described herein further comprise: a final concentration of about 11 wt% to about 16 wt% (e.g., about 11 wt% to about 15 wt%, about 14 wt%, about 13 wt%, or about 12 wt%, about 12 wt% to about 16 wt%, about 15 wt%, about 14 wt%, or about 13 wt%, about 13 wt% to about 16 wt%, about 15 wt%, or about 14 wt%, about 14 wt% to about 16 wt% or 15 wt%, or about 15 wt% to about 16 wt%) of an a-S1-casein protein (e.g., any a-S1-casein protein described herein); and/or alpha-S2-casein (e.g., any of the alpha-S2-casein proteins described herein) at a final concentration of about 2 wt% to about 5 wt% (e.g., about 2 wt% to about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, about 2.5 wt% to about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, about 3.5 wt%, or about 3.0 wt%, about 3.0 wt% to about 5.0 wt%, about 4.5 wt%, about 4.0 wt%, or about 3.5 wt%, about 3.5 wt% to about 5 wt%, about 4.5 wt%, or about 4.0 wt%, about 4.0 wt% to about 5.0 wt%, or about 4.5 wt% to about 5.0 wt%).

In some examples of any of the compositions described herein, the α -S1-casein protein may be a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, north american caprine (mountain goat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth α -S1-casein protein; and/or the alpha-S2-casein protein may be a bovine, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna or woolly mammoth alpha-S2-casein protein.

Some examples of any of the compositions described herein further include one or more (e.g., two or three) of serum albumin (e.g., any of the serum albumin proteins described herein), lactoferrin (e.g., any of the lactoferrin proteins described herein), and transferrin (e.g., any of the transferrins described herein). In some examples of any of the compositions described herein, the serum albumin can be bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly echis serum albumin; the lactoferrin may be a cow, a human, a sheep, a goat, a buffalo, a camel, a horse, a donkey, a lemur, a panda, a guinea pig, a squirrel, a bear, a macaque, a gorilla, a chimpanzee, a goat (mountain coat), a monkey, an ape, a cat, a dog, a wallaby (wallaby), a rat, a mouse, a elephant, a opossum, a rabbit, a whale, a baboon, a gibbon, a orangutan, a mandrill (mandrill), a pig, a wolf, a fox, a lion, a tiger, an echidna or a mammoth lactoferrin; and/or the transferrin can be a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby (wallaby), rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mangrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth transferrin.

In some examples of any of the compositions described herein, the composition further comprises one or more color balancing agents (e.g., any of the colorants described herein, e.g., beta-carotene or annatto).

Any of the compositions described herein can have a pH of about 6.2 to about 7.2 (e.g., about 6.2 to about 7.0, about 6.2 to about 6.8, about 6.2 to about 6.6, about 6.2 to about 6.4, about 6.4 to about 7.2, about 6.4 to about 7.0, about 6.4 to about 6.8, about 6.4 to about 6.6, about 6.6 to about 7.2, about 6.6 to about 7.0, about 6.6 to about 6.8, about 6.8 to about 7.2, about 6.8 to about 7.0, or about 7.0 to about 7.2).

In various embodiments, the milk protein component comprises about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 6% milk protein on a dry weight or total weight basis. In some embodiments, the composition may comprise from about 0.5% to about 2.5%, from about 1% to about 2%, from about 2% to about 3%, or from about 4% to about 10% protein, on a dry weight basis or a total weight basis. In particular embodiments, the composition may comprise from about 10% to about 15% protein, on a dry weight basis or a total weight basis.

A wide range of caseins, including caseins having substantial homology to wild type casein, variants, mutants of casein, are expressed and incorporated as components of milk proteins.

Dry composition

Also provided is a powder composition comprising: a final concentration of about 3.6 wt.% to about 5.4 wt.% (e.g., about 3.6 wt.% to about 5.2 wt.%, about 5.0 wt.%, about 4.8 wt.%, about 4.6 wt.%, about 4.4 wt.%, about 4.2 wt.%, about 4.0 wt.%, or about 3.8 wt.%, about 3.8 wt.% to about 5.4 wt.%, about 5.2 wt.%, about 5.0 wt.%, about 4.8 wt.%, about 4.6 wt.%, about 4.4 wt.%, about 4.2 wt.%, or about 4.0 wt.%, about 4.0 wt.% to about 5.4 wt.%, about 5.2 wt.%, about 5.0 wt.%, about 4.8 wt.%, about 4.6 wt.%, about 4.4 wt.%, or about 4.2 wt.%, about 4.2 wt.% to about 5.2 wt.%, about 5.0 wt.%, about 4.8 wt.%, about 4.6 wt.%, about 4.4 wt.%, or about 4.2 wt.%, or about 5.4 wt.% casein, about 4.2 wt.% to about 5.2 wt.%, about 4 wt.%, or about 4.4.2 wt.%, or about 5.4 wt.%, or about 4.2 wt.%, or about 5 wt.% casein, 4.2 wt.%, about 4 wt.%, about 4.2 wt.%, about 5.2 wt.%, or about 5 wt.%, or about 5.4 wt.%, or about 5 wt.% A substance (e.g., any of the α -casein proteins described herein); a final concentration of about 16.3 wt% to about 24.5 wt%, 16.3 wt% to about 22 wt%, about 20 wt%, or about 18 wt%; about 18 wt% to about 24.5 wt%, about 22 wt%, or about 20 wt%; about 20 wt% to about 24.5 wt% to about 22 wt%; or about 22 wt.% to about 24.5 wt.%) of a beta-casein protein (e.g., any of the beta-casein proteins described herein); a sweetener (e.g., any one or more of the sweeteners described herein) at a final concentration of about 35% to about 40% by weight (e.g., about 35% to about 39%, about 38%, about 37%, or about 36%, about 36% to about 40%, about 39%, about 38%, or about 37%, about 37% to about 40%, about 39%, or about 38%, about 38% to about 40%, or 39%, or about 39% to about 40%) by weight; a final concentration of about 25 wt.% to about 30 wt.% (e.g., about 25 wt.% to about 29 wt.%, about 28 wt.%, about 27 wt.%, or about 26 wt.%, about 26 wt.% to about 30 wt.%, about 29 wt.%, about 28 wt.%, or about 27 wt.%, about 27 wt.% to about 30 wt.%, about 29 wt.%, or about 28 wt.%, about 28 wt.% to about 30 wt.% or about 29 wt.%, or about 29 wt.% to about 30 wt.%) of one or more lipids (e.g., any one or more lipids described herein); ash (e.g., any ash described herein) at a final concentration of about 5 wt.% to about 7 wt.% (e.g., about 5 wt.% to about 6.5 wt.%, about 6.0 wt.%, or about 5.5 wt.%; about 5.5 wt.% to about 7.0 wt.%, about 6.5 wt.%, or about 6.0 wt.%; about 6.0 wt.% to about 7.0 wt.%, or about 6.5 wt.%; or about 6.5 wt.% to about 7.0 wt.%); and water at a final concentration of about 2 wt.% to about 5 wt.% (e.g., about 2 wt.% to about 4 wt.% or about 3 wt.%, about 3 wt.% to about 5 wt.% or about 4 wt.%, or about 4 wt.% to about 5 wt.%), wherein the kappa-casein protein is unglycosylated and/or has a non-mammalian glycosylation pattern, and/or the beta-casein protein is unglycosylated and/or has a non-mammalian glycosylation pattern.

Any of the powder compositions can contain any of the components described in any of the compositions described herein (e.g., any one or more of the color-imparting agents, alpha-S1-casein protein, alpha-S2-casein protein, alpha-lactalbumin protein, beta-lactoglobulin protein, lactoferrin, transferrin protein, and serum albumin protein, respectively, at any concentration described herein for each component).

Supplemented milk compositions

Also provided herein are compositions comprising: mammalian-produced milk or processed mammalian-produced milk; and one or more (e.g., two or three) of the following: kappa-casein proteins that are non-glycosylated or have a non-mammalian glycosylation pattern; a β -casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern; or micelles comprising kappa-casein protein which is non-glycosylated or has a non-mammalian glycosylation pattern and beta-casein protein which is non-glycosylated or has a non-mammalian glycosylation pattern.

In some examples, the composition includes mammal-produced milk or processed mammal-produced milk and kappa-casein protein that is not glycosylated or has a non-mammalian glycosylation pattern. In some examples, the composition comprises mammal-produced milk or processed mammal-produced milk and β -casein protein that is not glycosylated or has a non-mammalian glycosylation pattern. In other examples, the composition comprises mammal-produced milk or processed mammal-produced milk and micelles comprising kappa-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern and beta-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern.

In some examples, the final concentration of non-glycosylated or non-mammalian glycosylation pattern kappa-casein protein or non-glycosylated or non-mammalian glycosylation pattern beta-casein protein in the composition is: 0.02 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, about 0.4 wt%, about 0.2 wt%, or about 0.1 wt% (of the final composition); about 0.1 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, about 0.4 wt%, or about 0.2 wt%; about 0.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, or about 0.4 wt%; about 0.8 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, or about 1.0 wt%; about 1.0 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, or about 1.2 wt%; about 1.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, or about 1.4 wt%; about 1.4 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, or about 1.6 wt%; about 1.6 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, or about 1.8 wt%; about 1.8 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, or about 2.0 wt%; about 2.0 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, or about 2.2 wt%; about 2.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, or about 2.4 wt%; about 2.4 wt% to about 3.0 wt%, about 2.8 wt%, or about 2.6 wt%; about 2.6 wt% to about 3.0 wt% or about 2.8 wt%; or from about 2.8 wt% to about 3.0 wt%.

In some compositions, the final concentration of non-glycosylated and/or non-mammalian glycosylation pattern kappa-casein protein in the composition is from about 0.02 wt% to about 0.6 wt% (e.g., from about 0.02 wt% to about 0.5 wt%, from about 0.02 wt% to about 0.4 wt%, from about 0.02 wt% to about 0.3 wt%, from about 0.02 wt% to about 0.2 wt%, from about 0.02 wt% to about 0.1 wt%, from about 0.1 wt% to about 0.5 wt%, from about 0.1 wt%, to about 0.4 wt%, from about 0.1 wt% to about 0.3 wt%, from about 0.1 wt% to about 0.2 wt%, from about 0.2 wt% to about 0.5 wt%, from about 0.2 wt% to about 0.4 wt%, from about 0.2 wt% to about 0.3 wt%, from about 0.3 wt% to about 0.5 wt%, from about 0.4 wt%, or from about 0.3 wt%; and the final concentration of non-glycosylated and/or non-mammalian glycosylation pattern beta-casein protein in the composition is from about 0.02 wt.% to about 4.0 wt.%, about 3.8 wt.%, about 3.6 wt.%, about 3.4 wt.%, about 3.2 wt.%, about 3.0 wt.%, about 2.8 wt.%, about 2.6 wt.%, about 2.4 wt.%, about 2.2 wt.%, about 2.0 wt.%, about 1.8 wt.%, about 1.6 wt.%, about 1.4 wt.%, about 1.2 wt.%, about 1.0 wt.%, about 0.8 wt.%, about 0.6 wt.%, about 0.4 wt.%, or about 0.2 wt.%; about 0.2 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, or about 0.4 wt%; about 0.4 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, or about 0.6 wt%; about 0.6 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, or about 0.8 wt%; about 0.8 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, or about 1.0 wt%; about 1.0 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, or about 1.2 wt%; about 1.2 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, or about 1.4 wt%; about 1.4 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, or about 1.6 wt%; about 1.6 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, or about 1.8 wt%; about 1.8 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, or about 2.0 wt%; about 1.8 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, or about 2.0 wt%; about 2.0 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, or about 2.2 wt%; about 2.2 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, or about 2.4 wt%; about 2.4 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, about 2.8 wt%, or about 2.6 wt%; about 2.6 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, about 3.0 wt%, or about 2.8 wt%; about 2.8 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, about 3.2 wt%, or about 3.0 wt%; about 3.0 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, about 3.4 wt%, or about 3.2 wt%; about 3.2 wt% to about 4.0 wt%, about 3.8 wt%, about 3.6 wt%, or about 3.4 wt%; about 3.4 wt% to about 4.0 wt%, about 3.8 wt%, or about 3.6 wt%; about 3.6 wt% to about 4.0 wt% or about 3.8 wt%; or from about 3.8 wt% to about 4.0 wt%.

In some examples, the final concentration of micelles comprising non-glycosylated or non-mammalian glycosylation pattern kappa-casein protein and non-glycosylated or non-mammalian glycosylation pattern beta-casein protein in the composition is: 0.02 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, about 0.4 wt%, about 0.2 wt%, or about 0.1 wt% (of the final composition); about 0.1 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, about 0.4 wt%, or about 0.2 wt%; about 0.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, about 1.0 wt%, about 0.8 wt%, about 0.6 wt%, or about 0.4 wt%; about 0.8 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, about 1.2 wt%, or about 1.0 wt%; about 1.0 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, about 1.4 wt%, or about 1.2 wt%; about 1.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, about 1.6 wt%, or about 1.4 wt%; about 1.4 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, about 1.8 wt%, or about 1.6 wt%; about 1.6 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, about 2.0 wt%, or about 1.8 wt%; about 1.8 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, about 2.2 wt%, or about 2.0 wt%; about 2.0 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, about 2.4 wt%, or about 2.2 wt%; about 2.2 wt% to about 3.0 wt%, about 2.8 wt%, about 2.6 wt%, or about 2.4 wt%; about 2.4 wt% to about 3.0 wt%, about 2.8 wt%, or about 2.6 wt%; about 2.6 wt% to about 3.0 wt% or about 2.8 wt%; or from about 2.8 wt% to about 3.0 wt%.

Nucleic acids and vectors

Also provided are nucleic acids (e.g., vectors) comprising: a promoter (e.g., a yeast, bacterial, or mammalian promoter); a sequence of encoded signal sequences; a sequence encoding a milk protein (e.g., any of the exemplary sequences described herein); and a yeast termination sequence, wherein the promoter is operably linked to a signal sequence operably linked to a sequence encoding a milk protein and the termination sequence is operably linked to a sequence encoding a milk protein. In some examples of these nucleic acids, the promoter is a constitutive promoter or an inducible promoter. Non-limiting examples of promoters are described herein. Additional promoters useful in these nucleic acids are known in the art.

The signal sequence in any of the vectors described herein may be a signal sequence from the encoded milk protein or a different milk protein, or a signal sequence from a yeast mating factor (e.g., any alpha mating factor). In some examples, the encoded milk protein is selected from: beta-casein (e.g., any beta-casein protein described herein), kappa-casein (e.g., any kappa-casein protein described herein), alpha-S1-casein (e.g., any alpha-S1-casein protein described herein), alpha-S2-casein (e.g., any alpha-S2-casein protein described herein), alpha-lactalbumin (e.g., any alpha-lactalbumin protein described herein), beta-lactoglobulin (e.g., any beta-lactoglobulin protein described herein), lactoferrin (e.g., any lactoferricin described herein), or transferrin (e.g., any transferrin protein described herein). Additional signal sequences useful in the vectors of the invention are known in the art.

Any of the nucleic acids described herein can also include a bacterial origin of replication. Any of the nucleic acids described herein can also include a selectable marker (e.g., an antibiotic resistance gene). The sequences of bacterial origins of replication are known in the art. Non-limiting examples of antibiotic resistance genes are described herein. Additional examples of resistance genes are known in the art.

Non-limiting examples of termination sequences are described herein. Additional examples of termination sequences are known in the art.

Some embodiments of the nucleic acids provided herein further comprise: additional promoter sequences (e.g., any of the exemplary promoters described herein); a further sequence encoding a signal sequence (e.g., any of the exemplary signal sequences described herein); a sequence encoding an additional milk protein (e.g., any exemplary sequence encoding a milk protein described herein); and an additional yeast termination sequence (e.g., any of the exemplary yeast termination sequences described herein), wherein the additional promoter sequence is operably linked to the additional sequence encoding the signal sequence, the sequence encoding the signal sequence is operably linked to the sequence encoding the additional milk protein, and the sequence encoding the additional milk protein is operably linked to the additional yeast termination sequence. The additional milk protein may be, for example, beta-casein (e.g., any beta-casein protein described herein), kappa-casein (e.g., any kappa-casein protein described herein), alpha-S1-casein (e.g., any alpha-S1-casein protein described herein), alpha-S2-casein (e.g., any alpha-S2-casein protein described herein), an alpha-lactalbumin (e.g., an alpha-lactalbumin protein described herein), a beta-lactoglobulin (e.g., any beta-lactoglobulin protein described herein), a lactoferrin (e.g., any lactoferricin described herein), or a transferrin (e.g., any transferrin protein described herein). In some embodiments, the nucleic acid comprises a sequence encoding beta-casein and a sequence encoding kappa-casein. The promoter and the further promoter may be the same or different. The yeast termination sequence and the additional yeast termination sequence may be the same or different. The signal sequence and the further signal sequence may be the same or different.

The invention also includes vectors comprising an isolated DNA sequence encoding a casein or whey polypeptide and host cells comprising the vectors. The vector may further comprise an isolated DNA sequence comprising a nucleotide sequence encoding casein (wherein the nucleotide sequence is operably linked to a promoter), a nucleotide sequence encoding an alpha mating factor or variant thereof, a nucleotide sequence encoding a bacterial resistance marker and a transcription terminator. One or more suitable promoters for expressing the gene encoding casein or whey protein may be any promoter that is functional in the host cell and capable of inducing expression of the product encoded by the gene. Suitable promoters include, for example, P_LAC4-PBIT7, Ptac, Pgal, lambda PL, lambda PR, bla, spa, Adh, CYC, TDH3, ADH1, and CLB 1.

Introduction of nucleic acids into cells

Methods of introducing nucleic acids (e.g., any of the nucleic acids described herein) into a cell to produce a host cell are well known in the art. Non-limiting examples of techniques that can be used to introduce nucleic acids into cells include: calcium phosphate transfection, dendrimer transfection, lipofection (e.g., cationic lipofection), cationic polymer transfection, electroporation, cell extrusion, sonication, optical transfection, protoplast fusion, immunoperfection, hydrodynamic delivery, gene gun, magnetic transfection, and viral transduction.

Based on the knowledge that certain techniques in the art for introducing nucleic acids into cells are more effective for different types of host cells, one skilled in the art will be able to select one or more suitable techniques for introducing nucleic acids into cells. An exemplary method for introducing nucleic acids into yeast cells is described in Kawai et al,Bioeng.Bugs 1:395-403, 2010。

host cell

Also provided herein are host cells comprising any of the nucleic acids (e.g., vectors) described herein. In some examples, a nucleic acid described herein is stably integrated within the genome (e.g., chromosome) of the host cell. In other examples, the nucleic acids described herein are not stably integrated within the genome of the host cell.

In some embodiments, the host cell is a yeast strain or a bacterial strain. In some embodiments, the host cell may be, for example, a yeast strain selected from the group consisting of: kluyveromyces species, Pichia species, Saccharomyces species, Tetrahymena species, yarrowia species, Hansenula species, Brucella speciesBlastobotrys sp.), Candida species, Zygosaccharomyces species, and Debaryomyces species. Additional non-limiting examples of yeast strains that can be used as host cells are Kluyveromyces lactis, Kluyveromyces marxianus, Saccharomyces cerevisiae, and Pichia pastoris. Additional yeast strain species that can be used as host cells are known in the art.

In some examples, the host cell may be a protozoan, such as, for example, Tetrahymena thermophila,hegewischiTetrahymena, and Tetrahymena,hyperangularisTetrahymena, and Tetrahymena,malaccensisTetrahymena, darkling beetle, tetrahymena pyriformis and tetrahymena bulimia.

It is an object of the present invention to isolate milk protein fractions by recombinant expression thereof in any of the host cells provided herein.

Method for producing recombinant milk protein and method for producing micelle

Also provided is a method of producing a recombinant milk protein (e.g., any one or more of the milk proteins described herein) that is non-glycosylated or has a non-mammalian glycosylation pattern, comprising: culturing any of the host cells described herein in a culture medium under conditions sufficient to allow secretion of milk proteins that are non-glycosylated or have a non-mammalian glycosylation pattern; and harvesting the milk protein that is not glycosylated or has a non-mammalian glycosylation pattern from the culture medium. For these methodsSuitable media in (a) are known in the art. Culture conditions sufficient to allow secretion of milk proteins are also known in the art. The host cell used in these methods may be any of the host cells described herein. The host cell may comprise any of the nucleic acids described herein. The recombinant milk protein produced may be one or more of the following: beta-casein (e.g., any of the beta-casein proteins described herein), kappa-casein (e.g., any of the kappa-casein proteins described herein), alpha-S1-casein (e.g., any of the alpha-S1-casein described herein), alpha-S2-casein (e.g., any of the alpha-S2-casein described herein), alpha-lactalbumin (e.g., any of the alpha-lactalbumin proteins described herein), beta-lactoglobulin (e.g., any of the beta-lactoglobulin proteins described herein), lactoferrin (e.g., any of the lactoferrin proteins described herein), transferrin (e.g., any of the transferrin proteins described herein), and serum albumin (e.g., any of the serum albumin proteins described herein). Some of these methods also include isolating (e.g., purifying) the recombinant milk protein from the culture medium. Methods for isolating (e.g., purifying) recombinant milk proteins from liquids are well known in the art. Exemplary methods for isolating (e.g., purifying) recombinant milk proteins are described in Imafidon et al, Crit. Rev. Food Sci.Nutrition 37:663-669, 1997)。

Also provided are methods of producing micelles comprising non-glycosylated or non-mammalian glycosylation pattern beta-casein (e.g., any of the beta-casein proteins described herein) and non-glycosylated or non-mammalian glycosylation pattern kappa-casein (e.g., any of the kappa-casein proteins described herein), comprising: culturing any of the host cells described herein in a culture medium under conditions sufficient to allow the release of micelles from the host cell, wherein the host cell comprises a nucleic acid comprising a sequence encoding beta-casein and a sequence encoding kappa-casein; and harvesting the micelles from the culture medium. Suitable media for use in these methods are known in the art. The host cell used in these methods may be any of the host cells described herein. The host cell may comprise any of the nucleic acids described herein. The micelles produced may be any of the micelles described herein (and may have any of the physical characteristics of the micelles described herein). Some of these methods also include isolating (e.g., purifying) the micelles from the culture medium. Methods for separating (e.g., purifying) micelles from a liquid are well known in the art (e.g., ultracentrifugation).

Exemplary details of culturing yeast host cells are described in Idiris et al,Appl. Microbiol. Biotechnol.86:403-,Biotechnol. Bioprocess. Eng. 5:275-287, 2000; Zhu, Biotechnol. Adv. 30:1158-1170, 2012, Li et al,MAbs 2:466-477, 2010。

the present invention aims to express one or more different forms of casein for application in various types of dairy substitute products. Casein subunits such as α -s 1-casein, α -s 2-casein, β -casein and κ -casein differ by one or more amino acid changes. In certain embodiments, the methods and compositions include the incorporation of bovine casein such as α -s 1-casein, α -s 2-casein, β -casein, and κ -casein. In other embodiments, the methods and compositions include the incorporation of human casein such as beta-casein and kappa-casein. See U.S. patent No. 5,942,274. In an alternative embodiment, the casein is selected from one or more of the following sources, including but not limited to: cattle, humans, buffalos, camels, goats, sheep, horses, dolphins, whales, north american wild goats, and pigs.

Also provided are methods for producing a milk protein fraction, which may include, for example, using the plasmids or constructs of the invention as described in example 1. The method comprises preparing a plasmid of interest, inserting the plasmid into an appropriate host cell, culturing the host cell for a suitable time and under suitable conditions such that the protein of interest is expressed, and then purifying the protein.

Proteins can be separated based on their molecular weight, for example, by size exclusion chromatography, ultrafiltration through a membrane, or density centrifugation. In some embodiments, proteins may be separated based on their surface charge, for example, by isoelectric precipitation, anion exchange chromatography, or cation exchange chromatography. Proteins may also be isolated based on their solubility, for example by ammonium sulfate precipitation, isoelectric precipitation, surfactant, detergent or solvent extraction. Proteins can also be isolated by their affinity for another molecule using, for example, hydrophobic interaction chromatography, reactive dyes, or hydroxyapatite. Affinity chromatography may also include the use of antibodies with specific binding affinity for the protein, nickel NTA for His-tagged recombinant proteins, lectins that bind to sugar moieties on glycoproteins, or other molecules that specifically bind to the protein.

Typically, centrifugation at optimal pH gives purification efficiencies > 95%. The isoelectric points of native casein and whey proteins are known. Essentially, the pH of bovine alpha-s 1-casein is 4.91, the pH of bovine alpha-s 2-casein is 4.1, the pH of bovine beta-casein is 4.5, the pH of bovine kappa-casein is 4.1, the pH of bovine alpha-lactalbumin is 4.2, and the pH of bovine beta-lactoglobulin is 5.2. Recombinantly produced casein and whey may differ in their phosphate groups and carbohydrate groups. Other methods for protein purification include membrane filtration to remove any potential bacteria or contaminants, followed by lyophilization for protein isolation.

Preferably, the methods and compositions provide competitive production costs at or below $1,000/kg, $500/kg, $10/kg, $1.0/kg, $0.10/kg, $0.010/kg, or $0.0010/kg of the milk protein component. In a more preferred embodiment, the cost is less than $0.009, $0.007, $0.006, $0.005/kg of milk protein component.

Method of supplementing milk produced by a mammal

Also provided herein is a method of supplementing milk produced by a mammal, comprising: providing a mammal-produced milk or a processed mammal-produced milk; and mixing into the milk at least one of: a beta-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern (e.g., any of the beta-casein proteins described herein); a kappa-casein protein that is non-glycosylated or has a non-mammalian glycosylation pattern (e.g., any of the kappa-casein proteins described herein); and micelles comprising a β -casein protein that is unglycosylated or has a non-mammalian glycosylation pattern (e.g., any of the β -casein proteins described herein) and a κ -casein protein that is unglycosylated or has a non-mammalian glycosylation pattern (e.g., any of the κ -casein proteins described herein) (e.g., any of the micelles described herein).

One or more of beta-casein, kappa-casein and micelles may be mixed into milk to achieve any of the exemplary final concentrations of beta-casein protein, kappa-casein protein and micelles in the compositions described in the section herein referred to as "supplemented milk compositions". Methods of mixing are well known in the art. As will be appreciated by one skilled in the art, additional components described herein may also be mixed into the milk (e.g., without limitation, any of the components described herein).

Method for preparing composition

Also provided is a method of producing a composition comprising: sonicating a liquid comprising a protein mixture comprising beta-casein protein (e.g., any of the beta-casein proteins described herein) and kappa-casein protein (e.g., any of the kappa-casein proteins described herein), or comprising micelles comprising beta-casein protein (e.g., any of the beta-casein proteins described herein) and kappa-casein protein (e.g., any of the kappa-casein proteins described herein); mixing ash (e.g., any of the ashes described herein) into a liquid; adding to the liquid a mixture of one or more lipids (e.g., any one or more of the liquids described herein), one or more flavor compounds (e.g., any one or more of the flavor compounds described herein), and one or more color-balancing agents (e.g., any one or more of the color-balancing agents described herein), and sonicating the liquid; and adding one or more sweeteners (e.g., any one or more of the sweeteners described herein) to the liquid, thereby producing the composition.

In some examples of these methods, the beta-casein protein is non-glycosylated or has a non-mammalian glycosylation pattern, and/or the kappa-casein protein is non-glycosylated or has a non-mammalian glycosylation patternMammalian glycosylation patterns. In some examples of these methods, the ash comprises one or more of: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some examples of any of these methods, the added ash comprises CaCl₂、KH₂PO₄And sodium citrate (e.g., two or three).

In some examples of these methods, the one or more lipids comprise at least one (e.g., two, three, four, five, six, or seven) of: sunflower oil, coconut oil, tributyrin, mono-and diglycerides, free fatty acids and phospholipids. In some examples of these methods, the free fatty acid comprises at least one fatty acid selected from the group consisting of: butyric acid, caproic acid, caprylic acid and capric acid. In some examples of these methods, the phospholipid is soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid, or rapeseed lecithin phospholipid. In some embodiments of these methods, the flavor compounds comprise at least one (e.g., two, three, four, five, or six) flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone.

In some examples of these methods, the one or more color balancing agents is beta-carotene or annatto. In some embodiments of these methods, the one or more sweeteners are sugars (e.g., glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, or tagatose) or artificial sweeteners (e.g., stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osthol, betadine, sorbitol, thaumatin, xylitol, potassium acetaminosulfonate, advatame, alitame, aspartame-acesulfame, sodium cyclamate, dulcin, dulcoside dihydrochalcone, neotame, or P-4000).

The pH of the resulting composition can be about pH 6.2 to about pH 7.4 (e.g., about 6.2 to about 7.2; about 6.2 to about 7.0, about 6.2 to about 6.8, about 6.2 to about 6.6, about 6.2 to about 6.4, about 6.4 to about 7.2, about 6.4 to about 7.0, about 6.4 to about 6.8, about 6.4 to about 6.6, about 6.6 to about 7.2, about 6.6 to about 7.0, about 6.6 to about 6.8, about 6.8 to about 7.2, about 6.8 to about 7.0, or about 7.0 to about 7.2).

In any of these methods, the β -casein protein may be a bovine, human, ovine, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly mammoth β -casein protein; and/or the kappa-casein protein may be a bovine, human, ovine, caprine, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or mammoth kappa-casein protein.

In some embodiments of these methods, the protein mixture further comprises one or more proteins selected from the group consisting of: alpha-lactalbumin (e.g., any of the alpha-lactalbumin proteins described herein), beta-lactoglobulin (e.g., any of the beta-lactoglobulin proteins described herein), alpha-S1-casein (e.g., any of the alpha-S1-casein proteins described herein), alpha-S2-casein (e.g., any of the alpha-S2-casein proteins described herein), lactoferrin (e.g., any of the lactoferrin proteins described herein), transferrin (e.g., any of the transferrin proteins described herein), and serum albumin (e.g., any of the serum albumin proteins described herein).

As will be appreciated by one skilled in the art, the amount of each component used in these methods can be calculated in order to produce any of the compositions described herein.

Method for preparing butter, cheese, caseinate or yoghurt

Also provided herein is a method of preparing butter, cheese, caseinate, or yogurt comprising providing any of the compositions provided herein; and producing butter, cheese, caseinate, or yogurt using any of the compositions provided herein as starting materials.

Methods of preparing butter, cheese, caseinate, or yogurt are well known in the art. See, e.g., Scott, Cheesemaging Practice, Kluwer Academic/Plenum Publishers, New York, NY, 1998; U.S. patent No. 4,360,535 (which describes a method of making cream); u.s 285,878 (which describes a method of preparing butter).

Reagent kit

Also provided is a kit comprising: (a) a mixture of one or more milk proteins (e.g., any of the milk proteins described herein, including any one or more of the β -casein protein, κ -casein protein, α -S1-protein, α -S2 protein, α -lactalbumin protein, β -lactoglobulin protein, lactoferricin, transferrin protein, and serum albumin protein described herein), one or more lipids (e.g., any one or more of the lipids described herein), and one or more flavor compounds (e.g., any one or more of the flavor compounds described herein); and (b) a mixture of ash (e.g., any ash described herein) and at least one sweetener (e.g., any one or more sweeteners described herein). In some examples of these kits, the one or more milk proteins are cow, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, goat (mountain coat), monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboon, gibbon, orangutan, mandrill (mandrill), pig, wolf, fox, lion, tiger, echidna, or woolly ech milk proteins.

In some examples of these kits, the one or more fats are selected from: sunflower oil, coconut oil, tributyrin, mono-and diglycerides, free fatty acids and phospholipids. The fatty acids present in the kit may comprise at least one fatty acid selected from the group consisting of: butyric acid, caproic acid, caprylic acid and capric acid. The phospholipid in the kit can be soybean lecithin phospholipid, sunflower lecithin phospholipid, cotton lecithin phospholipid or rapeseed lecithin phospholipid.

The flavor compounds in the kit can include at least one flavor compound selected from the group consisting of: delta-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2, 3-pentanedione, gamma-undecalactone, and delta-undecalactone.

In some embodiments of the kit, the mixture in (a) further comprises one or more color balancing agents (e.g., any of the color balancing agents described herein, such as beta-carotene or annatto).

In some examples of the kit, the one or more sweeteners are sugars (e.g., glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, or tagatose) or artificial sweeteners (e.g., stevia, aspartame, cyclamate, saccharin, sucralose, mogroside, brazzein, curculin, erythritol, glycyrrhizin, inulin, isomalt, lactitol, mabinlin, maltitol, mannitol, miraculin, monatin, monellin, osthol, betadine, sorbitol, thaumatin, xylitol, potassium acetaminosulfonate, advantame, alitame, aspartame-acesulfame, sodium cyclamate, dulcin, dulcoside dihydrochalcone, neotame, or P-4000).

The kit may include ash comprising one or more of the following: calcium, phosphorus, potassium, sodium, citrate, and chloride. In some examples, the ash in the kit comprises CaCl₂、KH₂PO₄And sodium citrate (e.g.,two or three).

In some embodiments of the kit, the mixture in (a) is provided in a light-tight and air-tight package (e.g., a metal foil, such as aluminum foil), and/or the mixture in (b) is provided in an air-tight package (e.g., a sealed plastic bag).

Some examples of the kit further include instructions for preparing any of the compositions described herein.

Also provided herein are kits comprising at least one nucleic acid described herein.

Modulating flavor profile

Sensory impressions such as "feed-like", "bland" or "dirty" are described as flavour descriptions absorbed from the food ingested by the cattle and the surrounding odours. Others develop through microbial action due to the growth of a large number of bacteria. Chemical changes can also be made by enzymatic action, contact with metals such as copper, or exposure to sunlight or intense fluorescence. Quality control director is constantly striving to avoid off-flavors in milk and other dairy products. It is therefore an object of the present invention to reduce, eliminate or even mask the undesirable flavors and odors of various dairy products.

In certain preferred aspects of the invention, varying the fat content can alter the flavor and odor of various dairy substitute products. For example, increasing the butyric acid content may change the flavor profile of non-dairy cheese to resemble that of parma cheese. In other embodiments, modulating triglycerides such as caproic acid, capric acid and/or caprylic acid results in a flavor profile similar to goat cheese. Thus, adjusting the triglyceride with the ratio of the fatty acid components provides different flavor profiles that can be tailored to resemble those of various desired dairy products.

Similarly, the methods and compositions minimize one or more undesirable aromas by adjusting the various triglycerides incorporated into the dairy substitute product.

In certain aspects, this is mediated by the incorporation of synthetic short chain triglycerides in a desired combination in combination with a plant-based oil (e.g., sunflower oil). For example, a mixture of [ C18C 18C 6] and [ C18C 6C 18] provides a different flavor profile than a mixture of [ C18C 4C 4] and [ C18C 10C 10 ].

Dairy product substitute

A wide variety of dairy substitute products can be prepared using the methods and compositions of the present invention. Such products include, but are not limited to, milk, whole milk, buttermilk, skim milk, infant formula, condensed milk, dried milk, steamed milk, butter, clarified butter, cream, and various types of cheese.

Dairy substitute products may also be incorporated as dairy substitutes in a variety of food applications, including ice cream, frozen custard, frozen yogurt, cookies, cakes, cottage cheese, cream cheese, french fries, curds, and yogurt.

In certain aspects, the invention provides one or more subunits of casein selected from the group consisting of α -s 1-casein, α -s 2-casein, β -casein and κ -casein for use as a milk protein component in a dairy replacement product. A selected combination of casein subunits is used as the major or at least part of the milk protein component. In a preferred embodiment, the casein composition comprises casein subunits in such amounts that about 12-15g/L α -s 1-casein, about 3-4g/L α -s 2-casein, about 9-11g/L β -casein and about 2-4g/L κ -casein represent total casein in the synthetic dairy product.

In various embodiments, the casein composition may comprise about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 6% protein on a dry weight or total weight basis. In some embodiments, the casein composition may comprise about 0.5-2.5%, about 1-2%, about 2-3%, or about 4-10% casein protein, on a dry weight basis or a total weight basis. In a particular embodiment, the casein composition may comprise about 1.5-10% protein, on a dry weight basis or a total weight basis.

In certain aspects, the methods and compositions of the dairy substitute product are substantially free of one or more serum proteins. Serum proteins typically include proteins such as enzymes, hormones, growth factors, nutrient transporters, and disease resistance factors. In further embodiments, the methods and compositions of the dairy substitute product are substantially free of one or more immunoglobulins that may induce an undesired immune response.

In some embodiments, the whey composition may comprise, on a dry weight or total weight basis, about 0.001%, about 0.05%, about 0.1%, about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4% whey protein. In some embodiments, the composition may comprise from about 0.1% to about 1%, from about 1% to about 2%, from about 2% to about 3%, or from about 0.1% to about 2.3% protein, on a dry weight basis or a total weight basis. In particular embodiments, the composition may comprise from about 10% to about 15% protein, on a dry weight basis or a total weight basis.

In various embodiments, the carbohydrate is incorporated into a dairy replacement product. These carbohydrates provide a mild sweetness to the flavor profile of the product and also serve as a fast acting energy and nutrient source. Carbohydrates include, but are not limited to, sugars such as galactose, sucrose, glucose, fructose, and maltose. Sugar sources that are free of dairy products include, but are not limited to, sugar beets and other plants such as celery, basil, honey, cherry, corn, spinach, plum, kiwi and pea.

Lactose intolerance is common to many milk consumers. Thus, in a preferred embodiment, carbohydrates such as lactose are omitted from the dairy substitute composition. In preferred embodiments, the methods and compositions of the dairy substitute composition are substantially free of lactose.

In some embodiments, the carbohydrate composition may comprise about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5% carbohydrate on a dry weight basis or a total weight basis. In some embodiments, the composition may comprise from about 1% to about 3%, from about 2% to about 4%, or from about 10% to about 30% carbohydrate on a dry weight basis or a total weight basis. In particular embodiments, the composition may comprise from about 2% to about 5% carbohydrate on a dry weight basis or a total weight basis.

Ash is attributed to the structure and stability of casein micelles. Ash is very important to keep together an emulsion that is a milk or cream. Calcium and phosphate present in the ash interact with the fat globules and casein micelles to maintain an emulsified mixture.

The ash also affects sensory characteristics such as mouthfeel, body, and to some extent, milk flavor.

In some embodiments, the ash composition may comprise about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, or about 3% ash, on a dry weight or total weight basis. In some embodiments, the composition may comprise about 0.1-0.3%, about 0.5-0.7%, about 0.7-1%, or about 1-2% ash, on a dry weight or total weight basis. In particular embodiments, the composition may comprise from about 0.6% to about 0.8% protein, on a dry weight basis or a total weight basis.

Additional ingredients of various dairy products that are free of animal ingredients include vitamins, flavoring agents, natural or artificial sweeteners, colorants, salts, pH adjusters, binders, buffers, stabilizers, essential amino acids, anti-caking agents, anti-foaming agents, and mixtures thereof.

In some embodiments, the remaining ingredient composition may comprise about 0%, about 0.01%, about 0.1%, about 0.5%, about 1%, about 2%, about 3%, about 4%, or about 5% additives, on a dry weight basis or a total weight basis. In some embodiments, the composition may comprise from about 0.001% to about 0.01%, from about 0.01% to about 1%, from about 0.01% to about 2%, or from about 1% to about 5%, by dry weight or total weight, of an additive. In particular embodiments, the composition may comprise from about 0% to about 10% additives, on a dry weight basis or a total weight basis.

In some aspects, the present invention provides methods and compositions for a dairy substitute with fat comprising varying levels of triglyceride content. In a preferred embodiment, isolated triglycerides from various plant sources are incorporated with the milk protein component, carbohydrates and ash. It is an object of the present invention to modulate the percentage of fatty acids isolated in plants and transesterified in dairy substitute products to resemble the fatty acids found in natural dairy products and/or to develop novel flavor profiles with improved flavors not found in nature. In some embodiments, modulating specific short-to medium-chain fatty acids, including but not limited to butyric, capric, caprylic, caproic and lauric acids, provides desirable flavor profiles in dairy substitute products.

In some embodiments, the fat composition in the synthetic milk comprises about 0%, about 1%, about 2%, about 3%, about 3.5%, about 4% fat on a dry weight or total weight basis. In some embodiments, the composition may comprise about 1-2%, about 2-3%, about 3-4% fat on a dry weight basis or a total weight basis. In particular embodiments, the composition may comprise about 3-4% fat on a dry weight basis or a total weight basis. In alternative embodiments, the fat composition in the cream may comprise about 10%, about 20%, about 30%, about 40%, about 50% or even 60%. Preferably, the fat composition in the cream is generally from about 40 to about 50%.

In some aspects, short chain triglycerides are combined with longer chain oils to produce transesterified fatty acid esters. Preferably, the longer chain oil is selected from: sunflower, corn, olive, soybean, peanut, walnut, almond, sesame, cottonseed, canola, safflower, linseed, palm kernel, palm fruit, coconut, babassu, shea butter, mango butter, cocoa butter, wheat germ, and rice bran oil. More preferably, the longer chain oil comprises an engineered sunflower variety that overexpresses oleic acid by 400%.

Longer chain oils may also provide flavor characteristics, for example, reducing or even removing the overall flavor characteristics of the desired end product that is vibrant and soft.

In some embodiments, the fat component of the dairy substitute comprises selected triglycerides that are ester-exchanged to longer chain oils such as high oleic sunflower oil (example 2). It is expected that the same four short chain fatty acids may impart specific flavours, such as firmness and richness, to milk and derived products such as cheese. Various combinations of triglycerides and longer chain oils are incorporated to produce a number of different flavor profiles. In one embodiment, triglycerides with three oleic acids and in this case short chain triglycerides synthesized with one butyric acid, one caproic acid and one caprylic acid, yield the desired synthetic "milk fat" triglycerides. Additional embodiments include the incorporation of various short chain triglycerides to adjust slightly different flavor profiles, such as short chain triglycerides comprising caproic acid; short chain triglycerides comprising hexanoic acid and butyric acid; short chain triglycerides comprising hexanoic acid and decanoic acid. Thus, the methods and compositions provide various combinations of synthetic short chain triglycerides with sunflower oil triglycerides, resulting in different flavor profiles.

Synthetic milk

An exemplary embodiment of a synthetic milk formulation comprising the microbially-derived proteins of the present invention is illustrated in example 4. For example, the formulations of the invention incorporate all four subunits of bovine casein: alpha-s 1-casein, alpha-s 2-casein, beta-casein and kappa-casein and the two whey proteins alpha-lactalbumin and beta-lactoglobulin are the major milk protein components in the preparation. An exemplary synthetic milk formulation also comprises a plant-based interesterified fat as shown in figure 1. Additional components include carbohydrates and ash. The resulting milk substitute product exhibits characteristics that look, function, taste, smell, and feel like natural milk. As one of the key aspects of the invention, modifying the formulation of synthetic milk may exhibit different sensory impressions, such as flavour, by adjusting the oil content, i.e. the type of triglyceride added to the milk to mimic different flavours.

Such as Young W. Park, biological Components in Milk and Dairy Products, Technology&As described in Engineering, pp 60, 2009, sterols are a small fraction of the total lipid in milk, the main sterol is cholesterol (300 mg/100 g fat, which is equivalent to 10 mg/100 mL cow's milk) (Park et al, Small Rumin. Res.68:88-113, 2007). Goat milk has been shown to contain less cholesterol than other milks, but generally contains higher total fat than cow milk. See, Posati et al, 1976, Composition of foods, Agric. Handbook number 8-1, ARS, USDA, Washington, D.C., 1976, Jenness,J. Dairy Sci.63: 1605-. The cholesterol content is in different varietiesWith the majority of cholesterol in goat milk being in the free state, with only a small amount of 52 mg/100 g fat in the ester form. See, e.g., Arora et al, Ind. J. Dairy Sci. 29: 191.

In certain embodiments, the methods and compositions of the present invention provide synthetic dairy products with less or no cholesterol or the same cholesterol content as compared to dairy products by adjusting the oil content, i.e., the type of triglyceride. In other embodiments, the amount of saturated and unsaturated fats is also adjusted to at least less or the same amount of fat in the dairy substitute as compared to dairy milk. In a preferred embodiment, the synthetic dairy product of the invention has a very low saturated fat, but smells and tastes like milk. In a preferred embodiment of the invention, the long chain fatty acid is typically a saturated fatty acid in milk, rather than a monounsaturated acid, such as oleic acid.

The present invention may not require or at least minimize pasteurization, as each component may be separately sterilized prior to combination by the formulation method. In other embodiments of the invention, the synthetic dairy product of the invention may be subjected to pasteurization.

Homogenization is optional for the methods and compositions of the invention, as is the case with natural milk. When sold as a stand-alone liquid beverage, the synthetic dairy product of the invention may be sold in a homogeneous form.

Differences between the milk replacer of the present invention and milk include flavour, nutritional value and storage stability. The flavour may be adjusted to the desired sensory impression based on triglycerides and other natural or artificial flavours that may impart off-taste or clarity or different flavours, such as cow, goat, coconut, almond or soybean.

Synthetic cheese

In other aspects of the invention, methods and compositions are provided comprising one or more isolated milk protein components, fat, carbohydrates and ash to produce various types of cheese products. Typically, the cheese is made from the milk protein component of the invention. One or more sensory impressions are incorporated into the cheese product by adjusting the triglycerides. Thus, cheese with desired organoleptic characteristics of different appearance, aroma, taste and texture can be produced. For some cheese varieties, in addition to conditioning triglycerides, one or more bacteria are used in the cheese making process for fermentation, wherein fermentation products and by-products such as lactic acid, carbon dioxide, alcohols, aldehydes, and ketones are produced. Types of cheese include whey cheeses such as ricotta and mozzarella, semi-soft cheeses including havatia and munster, medium hard cheeses such as Swiss and jarsberg, hard cheeses such as Cheddar and soft ripe cheeses such as Brie and Camembert.

Synthetic cream

The cream substitute directly usable should preferably comprise about 50 to 90 wt% water, more preferably about 65 to 80 wt% water, wherein the base is dispersed in the water. The base for the replacement cream should advantageously contain (all percentages calculated using the total weight of the base as 100%) from about 22 to 87% by weight of carbohydrates (more preferably from about 30 to 64%), from about 12 to 70% by weight of particulate fat (most preferably from about 28 to 60%) and from about 0.4 to 8% by weight of the selected emulsifier or group thereof (most preferably from about 1 to 4%).

In a preferred embodiment, the product of the invention is stable in aqueous emulsions. As used herein, a dried liquid fat-containing non-dairy food product is considered "stable" when the following minimum criteria are met: reconstituted emulsion stability, whitening ability, oiling or oil separation, feathering precipitation. See U.S. patent No. 4,310,561.

Synthetic butter

Commercial butter contains 80-82% milk fat, 16-17% water and 1-2% milk solids other than fat (sometimes referred to as curd).

Advantages of the dairy substitute products or compositions provided herein

The desired advantages of the present invention are environmental properties such as 8 times energy efficiency, 260 times water efficiency compared to conventional dairy products. Other environmental advantages include less water usage than conventional milk production, which is estimated to be about 1000L/L, and reduced land usage for traditional milk production typically requires grazing, crop land, the ability to reduce 6000 billion kg of carbon dioxide emissions from traditional milk production per year. The present invention also provides for reduction or elimination of costs of feed, handling, labor, animals, and marketing. Preferably, the feed cost is substantially reduced by a factor of 8.

Food safety advantages include reduction or removal of antibiotic residues, heavy metals, bacteria, adulterants. Thus, certain aspects of the present invention provide bacteria-free, animal component-free milk that does not require pasteurization or cold transport, but has an extended shelf life and exhibits many characteristics, such as taste, appearance, handling, and mouthfeel characteristics, that are the same as, or at least very similar to, its traditional dairy counterpart. Preferably, the dairy substitute product is substantially free of bacteria, such as brucella (a), (b), (c), (d) and (d)Brucella) Campylobacter (I), (II)Camplyobacter) Listeria (R), (L) and (L)Listeria) Mycobacterium genus (A), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C), (B), (C)Mycobacterium) Salmonella genus (A), (B)Salmonella) Shigella (A), (B), (C)Shigella) Yersinia genus (Yersinia) Giardia (Giardia) (II)Giardia) And norovirus (noroviruses) and are therefore safer to eat. Additional advantages include minimal or no pasteurization and/or homogenization. More preferably, the dairy substitute is shelf-stable for production and dispensing for a relatively long period of time (e.g., at least three weeks, preferably longer). Even more preferably, the dairy substitute product has a lower environmental impact.

Several aspects of the invention are described below with reference to exemplary applications for illustration. It should be understood that numerous specific details, relationships, and methods are set forth to provide a full understanding of the invention. One skilled in the relevant art will readily recognize, however, that the invention can be practiced without one or more of the specific details or with other methods.

Examples

Example 1

Carrier

Protein sequence bovine alpha-S1 casein (U)niProt accession number P02662), bovine alpha-2 casein (UniProt accession number P02663), bovine beta-casein (UniProt accession number P02666), bovine kappa-casein (UniProt accession number P02668), bovine alpha-lactalbumin (UniProt accession number B6V3I5) and bovine beta-lactoglobulin (UniProt accession number P02754) were obtained at uniprot.org with the following changes: removing the 15 or 21-residue signal peptide from the N-terminus; adding to the 5' end of the DNAXhoI(CTC GAG) endonuclease recognition sequence and KEX endopeptidase recognition sequence (AAA AGA); and adding to the 3' end of the DNASalI(GTC GAC) endonuclease recognition sequence. Additional combined sequences were prepared by combining the sequences of the four caseins in the order shown above, each sequence was isolated with the following DNA fragments:

GGC TCA GGA TCA GGG TCG AAA AGA GGC TCA GGA TCA GGG TCG (SEQ ID NO:128)。

here, the non-underlined fragment codes (GS)₆The linker sequence serves for sufficient post-translational spacing and accessibility to the KEX protease, and the underlined segment encodes the KEX endopeptidase sequence of the post-translationally cleaved protein. As above, the entire cassette was flanked at the 5' endXhoIAnd flanked at the 3' end bySalIFor ligation into pKLAC2 (New England Biolabs, Beverly, Mass.). DNA was synthesized by Gen9, inc. (Cambridge, MA) or IDT (Coralville, IA). The plasmids used have, inter alia, a multiple cloning site, the Lac promoter, an acetamide-based reporter gene and an alpha-mating factor gene, which are used as fusion proteins for the secretion of foreign proteins.

Yeast transfection

Transfection of yeast was accomplished by thawing a 0.5mL tube of competent cells containing 25% glycerol on ice and adding 0.62mL of yeast transfection reagent. The mixture was then heated at 30 ℃ for 30 minutes and heat shocked at 37 ℃ for 1 hour. Cells were then pelleted at 7000rpm and washed twice with 1.0mL YPGal medium. The cell mixture was then transferred to a sterile culture tube and incubated at 30 ℃ for 3 hours with constant shaking at 300 rpm. The cell mixture was then transferred to a sterile 1.5mL microcentrifuge tube and the cells were pelleted at 7000rpm for 2 minutes and resuspended in 1mL sterile 1X PBS. 10, 50 and 100. mu.L of the cell suspension were placed in separate fresh sterile 1.5mL microcentrifuge tubes each containing 50. mu.L sterile deionized water. The tubes were mixed briefly and spread onto separate yeast carbon-based agar (YCB agar) plates containing 5mM acetamide for selection. The plates were then incubated in an inverted fashion at 30 ℃ for 4 days until colonies formed. 15 individual colonies were then streaked onto fresh YCB agar plates containing 5mM acetamide and incubated at 30 ℃ for 2 days.

DNA encoding alpha-lactalbumin and beta-lactoglobulin (two major lactalbumins) was designed internally and synthesized pre-determined from IDT and transfected into competent kluyveromyces lactis cells from the New England Biolabs kit (catalog No. E1000S) according to the protocol provided by the supplier.

High throughput transfectant selection

From each YCB agar plate, each of 30 plates was tested for successful integration of the vector plasmid once the colonies had grown sufficiently. This was followed by PCR analysis of each plate to test for specific cells with multiple vector integrants. Once isolated, the individual culture with the highest yield was used for scale-up culture. This process can be iterated with successively higher concentrations of selection pressure to force colonies to form higher copy numbers of our engineered plasmid.

Transfection events were performed 5 times and plated on 5 plates consisting of nitrogen-free yeast carbon-based medium. (thus, any observed growth on these plates would suggest successful uptake of the plasmid if the exogenous DNA itself were not taken up). Of these 5 plates, 100% showed positive growth. 30 individual colonies from 5 plates were selected for amplification culture and each was grown on individual YCB agar plates to produce homozygous culture plates to allow easy characterization and management. After a 3-day growth period, individual colonies from each plate were initially added to 10ml glass culture tubes containing 2ml YPGal medium to test protein expression. After a two-day growth period, cells were pelleted out and the supernatant was run on an SDS PAGE gel to check for protein expression. The strains that provided the best protein expression were grown up to 10ml, 100ml, 500ml and finally 1L culture vessels. 2 liters of culture from each whey protein was grown. About 1 gram of protein was collected from the total amount, indicating a non-optimal yield/productivity of 0.5 g/L.

Amplification culture in 1L Shake flask culture

Cultures were grown up and inoculated in 1L shake flasks at a split ratio of at least 1: 10. The inoculated flasks were grown in production medium without acetamide supplementation for 24 hours prior to inoculation. On the day of start of the fed batch production run, the reactor was charged to 90% of the target start volume and heated to the run temperature. Now, the temperature is set at 30 ℃ to save energy costs associated with heating the reactor. Additional parameters may be explored in the process optimization stage. When the reactor reached 30 ℃, the inoculum flask was added dropwise to the reaction vessel using a peristaltic pump. The reactor was maintained at the target pH using software provided by the supplier. Samples were taken from the reactor broth twice daily to quantify the glucose and electrolyte usage of the cells and as a double check of the pH and dissolved gas measurements of the reactor. After each measurement, a large dose of glucose was added to maintain the target glucose concentration of 10%, although this could also change in process development. When the cells reached maximum density, protein production was triggered by the addition of galactose, which elicited the promoter on our pKLAC2 plasmid. Galactose was replenished until the end of the run. The optimal run length can also be determined in the process development, but is set for 5 days of batch feed. After complete operation, the yeast cells are removed from the reactor and the protein is purified as discussed below.

Casein protein purification

The following casein proteins α -s1 casein, α -s2 casein and β -casein are inherently hydrophobic and precipitate out when secreted from yeast and contacted with water. Purification from the reactor medium involves collection of the protein from the surface of the culture medium followed by drying to isolate the pure protein. K-casein is inherently hydrophilic, and purification of k-casein involves a pH change of 4.6 in solution followed by centrifugation at 10,000 rcf. The combined casein cassette functions in the same way as kappa-casein.

Whey protein purification

α -lactalbumin: the isoelectric point of alpha-lactalbumin is 4.2. The solubility of the protein was lowest when the pH of the bioreactor media solution was reduced to 4.2. This knocked the protein out of solution and allowed to collect by centrifugation. Beta-lactoglobulin: similar to the purification of alpha-lactalbumin, the pH of the solution is lowered to 5.2, the isoelectric point of beta-lactoglobulin. This neutralises the charge of the protein and allows it to be collected by centrifugation at 14,000 rcf.

Protein purification

2L of the medium was centrifuged at 3,000g in a floor centrifuge to pellet the yeast cells. The precipitate was discarded and the supernatant was transferred to a new vessel, lowering the pH of the solution to 4.2 for alpha-lactalbumin and 5.2 for beta-lactoglobulin (figure 2A). This was followed by incubation of the supernatant in shake flasks at 35 ℃ for 30 minutes and centrifugation at 14,000g in a floor centrifuge to precipitate the protein mixture (FIG. 2B).

Protein characterization

After separation of the protein by centrifugation, the solid pellet and supernatant solution were run on a 14% SDA-PAGE gel to check for protein expression. Positive bands were observed at 14kDa and 18kDa (FIG. 3), which correlate with the size of bovine-derived alpha-lactalbumin and beta-lactoglobulin, respectively. Further characterization was performed to determine equivalence in primary sequence, glycosylation and phosphorylation.

Example 2

Synthesis of triglycerides

Milk fatty triglycerides were prepared by transesterifying short-chain triglycerides to high-oleic sunflower oil (oil from custom engineered sunflower variants expressing fatty acid esters in the following ratios as described in table 1):

table 1:

www.sunflowernsa.com/uploads/resources/51/warner_.pdf。

short chain triglyceride production

Short chain fatty acids, mainly responsible for the rich flavour in milk and cream, are molecules with an even number of carbons from 4 to 10 and are mixed in the following ratios as described in table 2:

table 2:

www.ncbi.nlm.nih.gov/pmc/articles/PMC2596709/pdf/FNR-52-1821.pdf。

table 3:

chain length	Name (R)	Mass fraction in mixture (%)
			4	Butyric acid/butyric acid	40
6	Hexanoic/caproic acid	26
			8	Octanoic/octanoic acids	11
10	Capric/capric acid	22

The scores in table 3 are based on the relative presence of these substances in bovine milk, but can be varied during process development in order to design better tasting products and to design milk of other species such as buffalo or goat. The short chain fatty acids in the mass ratios indicated above were combined with toluene, p-toluenesulfonic acid and glycerol in a Dean-Stark trap, which is commonly used for esterification reactions, to remove water produced in the condensation reaction. The reaction was carried out in a fume hood for several hours until a water level into the trap was observed that lasted for over 30 minutes. The vessel was allowed to cool and the mixture was removed from the reaction flask. The mixture was washed twice with 5% sodium carbonate solution and 5 times with pure water. Brine (10% aqueous NaCl) was added periodically to break the emulsion formed in the separatory funnel. The washed mixture of short chain triglycerides, water, toluene and impurities was dried in a rotary evaporator at 90 ℃ and 54mbar atmosphere for 1 hour until its excess proceeded well in order to minimize the chance of food contamination.

Transesterification

The short chain triglyceride mixture was combined with high oleic sunflower oil in a volume ratio of 1: 8. Sodium methoxide in an amount equal to 1% by mass of the oil mixture was added to catalyze the transesterification reaction, and the reaction vessel was heated to 65 ℃ and stirred continuously under an inert argon atmosphere for 6 hours. A 5% acetic acid mixture was added to quench the reaction, then the oil was washed 5 times with deionized water and dried in a rotary evaporator at > 90 ℃ for 1 hour. The finished milk fat is autoclaved to ensure sterility and is therefore suitable for use in milk or cream as described above.

Example 3

Milk preparation

One non-limiting formulation of a milk composition is described below.

Table 4:

components	% w/v range	Amount (g/L)
			Casein protein	1 – 10	10 – 100
Whey protein	0 – 1	0 – 10
			Plant-based milk fat	0 – 8	0 – 80 ml/L
Candy	0 – 5	0 – 50
			Ash content	0.1 – 1	1 – 10
Calcium carbonate	0.1 – 0.5	1 – L
			X (function additive)	0 - 1	0 – 10L

The milk formulation was achieved by the following procedure per 1 liter of milk according to table 4. 26 grams casein, 3.5 grams whey and 5 grams ash were combined and mixed thoroughly. 40mL of triglyceride was thawed and heated to 55 ℃. The protein mixture was slowly poured into the triglyceride and vortexed at high speed for 5 minutes. Meanwhile, 3.5 grams of whey and 24 grams of galactose were added to 850 mL of deionized water; the mixture was heated to 37 ℃. The triglyceride/protein/ash mixture was transferred to a Waring commercial blender and blended at low speed. Slowly pouring the whey/galactose/water mixture into a mixer; the lid is placed on the stirrer. The mixture was blended at high speed for ten minutes. Deionized water was added to a final volume of 1000 mL. The milk may optionally be homogenized using existing methods. The above scheme for cream or any milk preparation can be modified by varying the ratio of solids; however, our preliminary studies indicate that the presence of ash in the protein mixture and the separation of a significant proportion of whey can greatly affect the quality of the emulsion.

Example 4

Synthetic milk preparation

As preliminary conceptual evidence, to determine whether key components of milk can be reconstituted to form milk, dried food grade purified casein and research grade whey were purchased. Irish cream was obtained from a local source and purified fat was isolated therefrom by centrifuging the cream at 14,000 g. Finally, all minerals used were purchased from Sigma Aldrich.

The terms:

c-roux = a roux made by mixing casein protein and fat together while keeping the temperature of the mixture at 37 ℃.

W-batter = a batter made by mixing together whey protein and fat while keeping the temperature of the mixture at 37 ℃.

CW-roux = a roux prepared by first mixing casein and whey protein together in a mixture, adding fat and mixing at 37 ℃.

Table 5:

experiment of	Results
		Casein + fat + water	A light-yellow liquid, wherein the light-yellow liquid,bad taste, protein precipitation, and bad mouthfeel (watery).
Casein + water + fat	Pale yellow liquid, bad taste, protein precipitation, and bad mouthfeel (watery).
		(casein + fat) to make a batter. Oil paste and water	Yellowish liquid, even taste, and poor mouthfeel (watery). Low protein precipitation was observed.

The poor mouthfeel (e.g., watery) is assumed to be due to a lack of whey protein.

Table 6:

experiment of	Results
		Casein, whey, fat and water	Pale yellow-white liquid, bad taste, precipitated protein, and bad mouthfeel.
C-oil paste + whey + water	Light yellow-white liquid, average taste, low protein precipitation, and bad taste
		W-oil paste + whey + water	Pale yellow-white liquid, average taste, low protein precipitation, and bad mouthfeel.
CW-oil paste + water	Pale yellow-white liquid, average taste, and bad mouthfeel. Zero protein precipitated.

Assuming that the mouthfeel is not good because of undesired casein micelle formation, the addition of Ca will allow the micelles to form again.

Table 7:

experiment of	Results
		CW-oil paste + water + calcium phosphate (optimum amount of Ca is calculated by experiment and error)	White liquid and normal taste. Zero protein precipitated. Taste averaging

To improve the taste, different sugars were added to the above mixture at different concentrations.

Table 8:

candy	2.4%	3.0%	3.6%	4.2%	4.8%
						Glucose	Good effect	Is too sweet	Is too sweet	Is too sweet	Is too sweet
Galactose	Bland taste	Is excellent in	Average	Is excellent in	Is too sweet
						Sucrose	Not good	Not good	Not good	Not good	Not good
Maltose	Bland taste	Is excellent in	Is excellent in	Is too sweet	Is too sweet

All other ions found in bovine milk were incorporated to recreate the ionic environment found in nature.

Reference documents: r, Rosmaninho, L.F. Melo/Journal of Food Engineering 73 (2006) 379-387

Table 9:

reagent	Composition (mM)
		KH₂PO₄	11.60
Potassium citrate H₂O^a	3.7
		Sodium citrate 2H₂O	6.1
K₂SO₄	1.03
		K₂CO₃	2.17
KCL	8.0
		CaCL₂. 2H₂O	8.98

The end result is a bright white liquid, probably because the ionic environment holds together the solids present in the milk and increases the overall refractive index of the solution. The taste is excellent, but it has an average mouthfeel (e.g. a certain amount of chalkiness is observed in the liquid). The precise mineral composition as described in table 9 can provide excellent mouthfeel.

Milk fat synthesis

Synthetic milk fat is prepared by transesterifying short chain fatty acids present in the large chain fatty acids present in high oleic sunflower oil triglycerides. The four short chains used are:

40% C4: butyric acid. Are found in milk, particularly goat, sheep and buffalo, butter, pamah, and as products of anaerobic fermentation (including in the colon and as body odor). It has an unpleasant and pungent taste with a sweet aftertaste (similar to diethyl ether). Butyric acid is present in humans and is the main characteristic odor of human vomit.

26% C6: caproic acid. A colorless oily liquid, has a fat-like, cheese-like, waxy and odor similar to that of goats or other farm animals.

11% C8: octanoic acid. It is an oily liquid that is sparingly soluble in water, with a slightly unpleasant rancid odor and taste.

22% C10 capric acid. There are not too many flavours to mention and the longer the carbon chain you start getting less flavour. This is in coconut oil, so it is not as much milk fat flavor as it is.

Iterations included lauric acid (C12) as it was present at 2.9% of the total fatty acid content in cow milk (Pearl-Rogers, J.; Dieffenbacher, A.; Holm, J.V. (2001). "Lexicon of lipid Nutrition" (IUPAC Technical Report), "Pure and Applied Chemistry 73 (4): 685 and 744. doi:10.1351/pac 200173685.)

In one quarter of The amounts specified below, as per Yu et al, The modification an analysis of The volatile oil for The recipe mapping.J. Am. Oil Chem. SocThe following procedure described in 77:911 (2000):

a mixture of butyric, caproic, caprylic and capric acids (Sigma Chemical Co., St. Louis, MO), 21.42g p-toluenesulfonic acid (Sigma Chemical Co.), 2.305mol glycerol (Sigma Chemical Co.), and 458mL toluene (Fisher Scientific) in the same ratio [ see above ] as found in milk fat samples and a total of 7.26mol was refluxed with a Dean-Stark trap for 6 hours. The reaction was considered complete when water was no longer dripping into the trap. SCTG was washed once with 5% sodium carbonate solution and several times with water. The SCTG was then heated at 85 ℃ in a rotary evaporator to remove water and toluene.

SCTG from commercial and natural sources were transesterified with HOSO (Trisun 80, RBD; AC Humko, Memphis, TN) at a SCTG/HOSO ratio of 1:8.82 to produce fat with the same percentage of SCFA as milk fat. SCTG from commercial sources was also transesterified at a SCTG/HOSO ratio of 1:7.19 to produce fat with SCFA at a level equal to 120% in milk fat. Sodium methoxide (Aldrich Chemical Company, St. Louis, MO) was used as a catalyst at 0.5% by weight of total oil. The reaction was stirred at 65 ℃ under nitrogen for 6 hours. Next, 5% acetic acid (Fisher Scientific) was added to neutralize the catalyst, and then the oil was washed several times with distilled water and dried on a rotary evaporator at 90 ℃ for 30 minutes.

A pilot scale Continuous deodorant similar to that described by Smouse (Smouse, T.H., A Laboratory Continuous Deodorizer, for 8: 1176-1181 (1997)) was used to deodorize transesterified oils. The oil flow rate was 600 mL/h, the column temperature was 180 ℃, the pressure was 0.5 torr, and the steam rate was 12.6 mL/h. Each batch of deodorized oil was tasted to ensure flavor. The deodorized oil was stored at 4 ℃ until used to prepare cheese.

Example 5

Regulation of fatty acids

Sunflower oil triglycerides with three oleic acids are transesterified with four short chain fatty acids containing one butyric acid, one caproic acid and one caprylic acid as part of the fat composition in the mixture of synthetic milk products. Such an array or combination of fats is expected to result in a synthetic milk fat that provides its rich taste compared to natural dairy milk. The ability to control the composition of one or more triglycerides may enhance or alter the flavor profile of the synthetic dairy product. Thus, long and short chain matrices (matrix) can be generated in flavor profiles, including but not limited to a variety of aromatic compounds associated with the sensory impressions buttery, nutty, sweet, sour, fruity, floral, bitter, woody, earthy, beany, spicy, metallic, sweet, musty, oily and vinegar. Furthermore, an increase in texture such as creaminess, an increase in melting properties or resistance and an increase in stretching capacity can be shown relative to corresponding dairy products.

Example 6 recombinant production of milk proteins

Alpha-lactalbumin, beta-lactoglobulin, alpha-S1-casein, alpha-S2-casein, beta-casein and kappa-casein were produced in recombinant yeast strains (Pichia pastoris). Since glycosylases in yeast are distinct from mammalian cells, proteins produced by yeast will be non-glucosylated or have a non-mammalian glycosylation pattern. The protein produced may be used as a component in any of the compositions described herein.

Plasmids

Plasmids were constructed for expression of each protein. Each plasmid comprises the following components: inducible (e.g., AOX1 promoter) or constitutive (GAP promoter or PGK promoter) promoters for each protein expressed; a sequence encoding a signal peptide for each protein, derived from the natural bovine protein sequence or from the yeast protein sequence (α mating factor or OST 1); a sequence encoding a milk protein to be expressed; yeast transcription terminator sequences for each protein expressed (e.g., AOX1, AOD, or CYC 1); a bacterial origin of replication from pUC19, which enables replication in e.coli; and a selectable marker cassette (e.g., kanR or zeocin r) that enables selection with antibiotics in bacteria and yeast.

The different plasmids used to produce the different proteins are listed in table 10 below.

TABLE 10 expression plasmids (SEQ ID NO)

¹159 (synthetic) SEQ ID NO

These plasmids were then integrated into wild-type pichia pastoris for expression. Protein production was detected by SDS-PAGE, ELISA and Western blotting.

Alpha-lactalbumin

Strain construction

Three plasmids were generated and expression of bovine alpha-lactalbumin (bvLAA) was placed in the methanol-inducible promoter P_AOX1Under the control of (a), which has a native LAA signal peptide (pJAG-nat-LAA), a full-length α mating factor signal peptide (pJAG-aMF-LAA) or an OST1 signal peptide (pJAG-OST-LAA).

Before transformation, 20. mu.g of each plasmid was linearized by digestion with the restriction enzyme SacI. The digested plasmid was then concentrated by ethanol precipitation and resuspended in 10. mu.l of distilled water.

Competent pichia pastoris cells were prepared as follows: a culture of Pichia pastoris was grown to log phase (OD 600-1.0) in YPD medium (10g/L yeast extract, 20g/L peptone, 20g/L glucose). A1.5 mL aliquot was collected by centrifugation and resuspended in 1mL of a 1:1 mixture of YPD +20 mM HEPES (pH 8):1M lithium acetate. After addition of 10. mu.L of 1M dithiothreitol, the cells were incubated for 15 minutes at 30 ℃ in a shaker at 300 rpm. Cells were pelleted by centrifugation and washed three times in 1mL of ice-cold 1M sorbitol. After the final wash, the cells were resuspended in 50 μ L of 1M sorbitol.

Cells were combined with linearized plasmid DNA in a cooled 2mm electroporation cuvette and pulsed at 1.5kV (25. mu.F, 200. omega.). The cells were transferred to a culture tube with 200. mu.L of cold 1:1 YPD: 1M sorbitol and allowed to recover at 30 ℃ (300rpm) for 2 hours. Finally, cells were plated on YPD agar plates containing bleomycin and grown at 30 ℃ for two days.

Protein expression

Colonies were picked from agar plates and grown in 750 μ L BMD1% (0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 1.1% glucose) at 30 ℃ at 300 rpm. After 48 hours, 750 μ L of BMM2(0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 1% methanol) was inoculated using 900 μ L of culture. After 24 hours, 150 μ L of BMM10 (BMM10:0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 5% methanol) and the sample was harvested for analysis a further day later.

Analysis of

Protein expression was analyzed in samples of cultures centrifuged to remove cell mass. The clarified supernatant was then assessed by SDS-PAGE, ELISA and Western blotting.

To visualize total protein via SDS-PAGE, cell-free supernatants were treated with SDS-PAGE sample buffer, boiled, and run on 10% polyacrylamide gels. The gel was stained with SYPRO Ruby stain (Life Technologies). The resulting gel showed secretion of a-lactalbumin to occur using OST1 or the native lactalbumin signal peptide (figure 4).

To measure protein titers via ELISA, 25 μ Ι _ of each sample was placed in a half-zone 96-well microtiter plate and allowed to bind overnight at 4 ℃. After sample removal, the binding surface was blocked by filling each well with 1% (w/v) Bovine Serum Albumin (BSA) dissolved in Tris buffered saline (50 mM Tris, pH 7.6, 150 mM NaCl) and incubating at room temperature for 1 hour. The samples were then incubated in primary antibody diluted in 1% BSA/TBS + 0.1% (v/v) Tween-20 for 1.5 hours. After three washes in TBS + Tween, the samples were incubated with horseradish peroxidase (HRP) -conjugated secondary antibodies for another hour. After three final washes in TBS + Tween, chromogenic substrate (TMB single solution, Life Technologies) was added and the absorbance at 650nm was measured. The data obtained showed that alpha-lactalbumin was secreted using the native alpha-lactalbumin signal peptide or the OST1 signal peptide (figure 5).

To analyze samples via western blot, one volume of sample was combined with an equal volume of SDS-PAGE sample buffer and run on a 10% polyacrylamide gel. Proteins were transferred to nitrocellulose membranes, which were blocked by treatment with 1% BSA/TBS for 1 hour. After 1.5 h incubation with primary antibody diluted in 1% BSA/TBS + Tween, the blot was washed three times in TBS + Tween. The blot was then incubated with a secondary antibody conjugated to horseradish peroxidase (HRP) for an additional hour. After three final washes in TBS + Tween, chromogenic substrate was added (1 step Ultra TMB blotting solution, Thermo Fisher). After staining was complete, the blot was washed in distilled water.

Beta-lactoglobulin

Strain construction

Three plasmids were assembled, and expression of bovine beta-lactoglobulin (bvLGB) was placed in the methanol-inducible promoter (P in pLH 37)_AOX1) Or one of two constitutive promoters (P in pLH 44)_GAPOr P in pLH45_PGK) Under the control of (3).

Before transformation, 20 μ g of pLH37 was linearized by digestion with the restriction enzyme SacI. Equal amounts of pLH44 and pLH45 were linearized with the enzyme ApaLI. The digested plasmid was then concentrated by ethanol precipitation and resuspended in 10. mu.l of distilled water.

Protein expression

To evaluate expression in clones transformed with a plasmid containing a methanol inducible promoter (pLH37), each clone was grown in 750 μ L BMD1% (0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 1.1% glucose) at 30 ℃ at 300 rpm. After 48 hours, 750 μ L of BMM2(0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 1% methanol) was inoculated using 900 μ L of culture. After 24 hours, 150 μ L of BMM10 (BMM10:0.2M potassium phosphate buffer, 13.4g/L yeast nitrogen source, 0.4mg/ml biotin, 5% methanol) and the sample was harvested for analysis a further day later.

To assess expression in clones transformed with plasmids supporting constitutive expression (pLH44 or pLH45), each clone was grown overnight in PG medium (20g/L peptone, 2% glycerol) at 30 ℃ with shaking at 300 rpm. Cultures were diluted 1:10 in minimal sulfate medium:

glucose 20g/L
	Calcium chloride (CaCl2) 1 g/L
Sodium phosphate (Na2PO4) 24 g/L
	Potassium sulfate (K2SO4) 18.2 g/L
Magnesium sulfate (MgSO4-7H2O) 14.9 g/L
	Ammonium sulfate (NH4)2SO 49 g/L
EDTA (ethylene diamine tetraacetic acid) 65.25 mg-L
	FeSO4-7H2O (iron sulfate heptahydrate) 12.18 g/L
ZnSO4-7H2O (Zinc sulfate heptahydrate) 25.0125 g/L
	CaCl2-2H2O (calcium chloride dihydrate) 12.615 g/L
CuSO4-5H2O (copper sulfate pentahydrate) 2.175 g/L
	NaMoO4-2H2O (sodium molybdate dihydrate) 2.088 g/L
CoCl2-6H2O (cobalt chloride hexahydrate) 2.0445 g/L
	MnCl2-4H2O (manganese chloride tetrahydrate) 1.392 g/L
Biotin 0.2175 g/L

After 48 hours, samples were collected for analysis.

Analysis of

Protein expression was analyzed in samples of cultures centrifuged to remove cell mass. The clarified supernatant was then evaluated by ELISA and western blotting.

Bovine casein

Dual expression plasmids were constructed to support the expression of α -S1-casein and β -casein in one plasmid and α -S2-casein and κ -casein in the other plasmid. These pairs were chosen because alpha-S1: α -S2: beta: kappa molar ratio of about 5.5: 1.5: 4.0: 1.5; it is therefore desirable to have similar copy numbers of α -S1-casein and β -casein, as well as similar copy numbers of α -S2-casein and κ -casein.

Beta-casein and alpha-S2-casein were placed under the control of the constitutive PGAP promoter in their respective plasmids, whereas alpha-S1-casein and kappa-casein were placed under the control of the constitutive PPGK promoter.

To introduce the protein into the secretory pathway, the protein with its native signal peptide (pLH46 and pLH47) or OST1 signal peptide (pLH48 and pLH49) is expressed. Furthermore, plasmids were prepared in which one protein with its native signal peptide was expressed and another protein with the signal peptide of OST1 was expressed:

pLH0050 OST 1-beta, native-alpha-S1

pLH0051 native-. beta.OST 1-. alpha. -S1

pLH0054 OST 1-alpha-S2, natural-kappa

pLH0055 native- α -S2, OST1- κ.

To generate strains expressing all four caseins, yeast cells were first transformed with plasmids encoding β -casein and α -S1-casein. Before transformation, 20. mu.g of each plasmid was linearized with the enzyme ApaLI. The digested plasmid was then concentrated by ethanol precipitation and resuspended in 10. mu.l of distilled water.

Cells were combined with linearized plasmid DNA in a cooled 2mm electroporation cuvette and pulsed at 1.5kV (25. mu.F, 200. omega.). The cells were transferred to a culture tube with 200. mu.L of cold 1:1 YPD: 1M sorbitol and allowed to recover at 30 ℃ (300rpm) for 2 hours. Finally, cells were plated on PG agar (20g/L peptone, 2% (v/v) glycerol, 2% agar) plates containing bleomycin and grown at 30 ℃ for two days.

Six clones from β + α S1 plates were then grown in culture and made available for DNA uptake using the procedure described above. They were then transformed with the linearized α S2+ κ plasmid and grown on PG plates containing G418 at 30 ℃ for two days.

Expression of

To assess the production of bovine casein protein, 5 clones expressing casein and a wild-type yeast negative control were grown overnight in PG medium (20g/L peptone, 2% glycerol) at 30 ℃ with shaking at 300 rpm. All five casein expressing clones expressed alpha 2 and kappa-casein with the corresponding native casein signal peptide. Clones sLH115, 116, 117, and 118 expressed beta-casein and alpha-S1-casein with respective native signal peptides; clone sLH122 expressed beta-casein and alpha-S1-casein with OST1 signal peptide. Cultures were diluted 1:10 in minimal sulfate medium:

Glucose 20 g/L
	Calcium chloride (CaCl2) 1 g/L
Sodium phosphate (Na2PO4) 24 g/L
	Potassium sulfate (K2SO4) 18.2 g/L
Magnesium sulfate (MgSO4-7H2O) 14.9 g/L
	Ammonium sulfate (NH4)2SO 49 g/L
EDTA (ethylene diamine tetraacetic acid) 65.25mg/L
	FeSO4-7H2O (iron sulfate heptahydrate) 12.18 g/L
ZnSO4-7H2O (Zinc sulfate heptahydrate) 25.0125 g/L
	CaCl2-2H2O (calcium chloride dihydrate) 12.615 g/L
CuSO4-5H2O (copper sulfate pentahydrate) 2.175 g/L
	NaMoO4-2H2O (sodium molybdate dihydrate) 2.088 g/L
CoCl2-6H2O (cobalt chloride hexahydrate) 2.0445 g/L
	MnCl2-4H2O (manganese chloride tetrahydrate) 1.392 g/L
Biotin 0.2175 g/L

After 48 hours, samples were collected for analysis.

Analysis of

Protein expression was analyzed in samples of cultures centrifuged to remove cell mass. The clarified supernatant was then evaluated by ELISA and western blotting.

To measure protein titers via ELISA, 25 μ Ι _ of each sample was placed in a half-zone 96-well microtiter plate and allowed to bind overnight at 4 ℃. After sample removal, the binding surface was blocked by filling each well with 1% (w/v) Bovine Serum Albumin (BSA) dissolved in Tris buffered saline (50 mM Tris, pH 7.6, 150 mM NaCl) and incubating at room temperature for 1 hour. The samples were then incubated in primary antibody diluted in 1% BSA/TBS + 0.1% (v/v) Tween-20 for 1.5 hours. After three washes in TBS + Tween, the samples were incubated with horseradish peroxidase (HRP) -conjugated secondary antibodies for another hour. After three final washes in TBS + Tween, chromogenic substrate (TMB single solution, Life Technologies) was added and the absorbance at 650nm was measured. ELISA data showed that different yeast strains can secrete α -S1 casein and β -casein into the culture medium (fig. 8).

The data in this example show that the different expression vectors described herein can be used to generate transgenic yeast strains that secrete different milk proteins.

Example 7 Process for preparing a composition

The exemplary compositions described herein were generated using the specific methods described below. A schematic of this process is shown in fig. 9.

For the preparation of dairy products, laboratory equipment such as mixers, stir plates and sonicators are used. For large scale production, standard fluid milk processing equipment should be used.

As shown in fig. 9, there are three main components used in the method of preparing the composition. These steps include:

A. Preparation of protein solutions

B. Preparation of oil mixtures

C. Reconstitution of milk solids.

In step a, powdered micellar casein and whey protein are mixed and blended (step 1), followed by mixing with Deionized (DI) water (step 2) to obtain a protein solution 1. Typically, this contained 2.8% powdered micellar casein, 0.7% powdered whey protein and 85.5% water in the solution. The mixing vessel was covered to prevent moisture evaporation. The mixing is carried out by a mixer, a stir plate or an ultrasonic instrument for a sufficient period of time (about 30 minutes). This mixing time ensures that all the protein is dispersed in the water. The mixing speed has been optimized to be moderate, which provides sufficient force to disperse the proteins and avoid entrapment of air in the solution. The water content can be adjusted according to the amount of other components.

In step 3, CaCl₂、KH₂PO₄And sodium citrate in water are a mineral source for producing mineral characteristics similar to natural cow's milk. In a typical case, CaCl₂The concentration of the solution is 0.1 g/mL, KH₂PO₄0.27 g/mL and the sodium citrate solution is 0.21 g/mL sodium citrate. Is used for preparing KH₂PO₄The water with the sodium citrate solution is usually warm to ensure KH ₂PO₄Is completely dissolved. During the mixing of the protein solution 1, 0.015% CaCl was slowly added₂(step 4). CaCl used₂The volume of the solution is determined by the required CaCl₂The weight percentage of (A) is adjusted. Mixing for about 30 minutes to allow the protein to mix with the Ca²⁺The ions are fully interacting. Subsequently, 0.27% KH was added₂PO₄And 0.21% sodium citrate were divided into 5 parts, and each part was slowly added to the mixed solution at intervals of 5 to 10 minutes (step 5). Adding 0.085% CaCl₂Divided into 4 portions, and the portions are slowly added to the mixed solution at intervals of 4 to 10 minutes (step 6). Mixing is continued for at least 30 minutes, preferably 1-2 hours, to obtain a protein solution 2.

In method B, low speed mixing is sufficient to achieve uniform mixing of the different oil components. The following percentages of each component used to prepare oil blend 1 are based on total oil blend 1 weight. Initially, 65% sunflower oil, 29% coconut oil and 2% tributyrin were mixed together to form an oil base (step 7). Sunflower oil and coconut oil are deodorized to prevent unwanted odors. The combination of sunflower oil, coconut oil and tributyrin may mimic the fatty acid profile of natural milk. The oil-based ingredients and their contents can be adjusted according to different needs (different types of products). An aroma mixture is prepared by mixing different aroma components in sunflower oil (step 8). Compounds for simulating aroma include, but are not limited to, ethyl butyrate, delta-decalactone, 2-furylmethyl ketone, 2, 3-pentanedione, gamma-undecalactone, delta-undecalactone, acetoin, furfuryl alcohol, furfural, 2-methylfurfural, and 2-methylpyrazine. The content thereof can be adjusted by various applications and preferences. Oil blend 1 was prepared by adding 2.5% monoglyceride and diglyceride, 0.6% free fatty acid, 0.5% phospholipid, and 0.4% fragrance blend to mix (step 9). In a typical case, the free fatty acids contain 0.15% butyric acid and 0.45% caproic acid. Soy lecithin was used as the phospholipid source. Soy lecithin is readily available and inexpensive. A solution of beta-carotene was prepared in sunflower oil at a concentration of 0.5 mg/g (step 10). The 4% oil mixture 1 and the 0.06% beta-carotene solution were mixed together to obtain oil mixture 2 (step 11). The amount of beta-carotene used was adjusted to achieve different color levels of the milk. The amount of oil mixture 1 may also be adjusted for different dairy applications.

In method C, oil mixture 2 is slowly added to protein solution 2 and mixed well to prepare product mixture 1 (step 12). Mixing may be performed by a mixer or sonicator. In a typical case, the oil mixture 2 and the protein solution 2 are mixed at a moderate to high speed to ensure that the oil is uniformly dispersed in the aqueous solution. Subsequently, sonication is applied to break down the oil globules into smaller sizes, which results in an increase in their stability in solution. It is necessary to prevent air bubbles from being trapped in the solution during mixing. Oil mixture 2 was stirred into the aqueous solution using a mixing time of at least 20 minutes and allowed to disperse thoroughly. A 4% maltose solution was added to product mixture 1 and mixing was continued for another 30 minutes to give product mixture 2 (step 13). The sweetness level can be adjusted by the sugar content according to different applications. The source of the sugar may also be adjusted as desired. Additional DI water may be required to bring the final total weight to 100%.

Intensive homogenization, pasteurization and sterilization are not included in the process. However, for scale-up culture production, it is necessary to apply these steps to prepare the product mixture in method C.

Apparatus for use

A mixer: IKA-Labortechnik RW16 Basic, speed level (4-6)

A tip ultrasonic instrument: qsonica model CL-188, amplitude 70%

Water bath ultrasonic instrument: bransonic model 1510R-MT.

Example 8 exemplary formulations

Exemplary formulation compositions having similar taste and texture characteristics as whole milk, cream, high protein milk, fat free milk, and sugar free milk are provided in tables 11-15 below.

The compositions listed in tables 11-15 were prepared by making the necessary modifications to the method described in example 7, as would be understood in the art.

Table 11. composition sample whole milk.

TABLE 12 cream compositions

Table 13. composition-like protein-rich milk.

TABLE 14. non-fat milks as compositions

Table 15. composition sample sugar free milk.

Example 9 exemplary compositions

An exemplary composition prepared by the presently described method is shown in fig. 10. The composition in fig. 10 has an appearance (color), viscosity, foamability, flavor, and nutritional value similar to milk produced by mammals. The composition shown in figure 10 comprises a protein of mammalian origin.

Sequence listing

<110> Pandya, Ryan

Gandhi, Perumal

<120> casein-containing composition and method for producing the same

<130> 42233-0002WO1

<150> 62/040,393

<151> 2014-08-21

<160> 159

<170> FastSEQ for Windows Version 4.0

<210> 1

<211> 190

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 1

Met Met Lys Ser Phe Phe Leu Val Val Thr Ile Leu Ala Leu Thr Leu

1 5 10 15

Pro Phe Leu Gly Ala Gln Glu Gln Asn Gln Glu Gln Pro Ile Arg Cys

20 25 30

Glu Lys Asp Glu Arg Phe Phe Ser Asp Lys Ile Ala Lys Tyr Ile Pro

35 40 45

Ile Gln Tyr Val Leu Ser Arg Tyr Pro Ser Tyr Gly Leu Asn Tyr Tyr

50 55 60

Gln Gln Lys Pro Val Ala Leu Ile Asn Asn Gln Phe Leu Pro Tyr Pro

65 70 75 80

Tyr Tyr Ala Lys Pro Ala Ala Val Arg Ser Pro Ala Gln Ile Leu Gln

85 90 95

Trp Gln Val Leu Ser Asn Thr Val Pro Ala Lys Ser Cys Gln Ala Gln

100 105 110

Pro Thr Thr Met Ala Arg His Pro His Pro His Leu Ser Phe Met Ala

115 120 125

Ile Pro Pro Lys Lys Asn Gln Asp Lys Thr Glu Ile Pro Thr Ile Asn

130 135 140

Thr Ile Ala Ser Gly Glu Pro Thr Ser Thr Pro Thr Ile Glu Ala Val

145 150 155 160

Glu Ser Thr Val Ala Thr Leu Glu Ala Ser Pro Glu Val Ile Glu Ser

165 170 175

Pro Pro Glu Ile Asn Thr Val Gln Val Thr Ser Thr Ala Val

180 185 190

<210> 2

<211> 573

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 2

atgatgaaga gttttttcct agttgtgact atcctagcat taaccctgcc atttttgggt 60

gcccaagagc aaaaccaaga acaaccaata cgctgtgaga aagatgaaag attcttcagt 120

gacaaaatag ccaaatatat cccaatccag tatgtgctga gtaggtatcc tagttatgga 180

ctcaattact accaacagaa accagttgca ctaattaata atcaatttct gccataccca 240

tattatgcaa agccagctgc agttaggtca cctgcccaaa ttcttcaatg gcaagttttg 300

tcaaatactg tgcctgccaa gtcctgccaa gcccagccaa ccaccatggc acgtcaccca 360

cacccacatt tatcatttat ggccattcca ccaaagaaaa atcaggataa aacagaaatc 420

cctaccatca ataccattgc tagtggtgag cctacaagta cacctaccat cgaagcagta 480

gagagcactg tagctactct agaagcttct ccagaagtta ctgagagccc acctgagatc 540

aacacagtcc aagttacttc aaccgcggtc taa 573

<210> 3

<211> 192

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 3

Met Met Lys Ser Phe Phe Leu Val Val Thr Ile Leu Ala Leu Thr Leu

1 5 10 15

Pro Phe Leu Gly Ala Gln Glu Gln Asn Gln Glu Gln Pro Ile Cys Cys

20 25 30

Glu Lys Asp Glu Arg Phe Phe Asp Asp Lys Ile Ala Lys Tyr Ile Pro

35 40 45

Ile Gln Tyr Val Leu Ser Arg Tyr Pro Ser Tyr Gly Leu Asn Tyr Tyr

50 55 60

Gln Gln Arg Pro Val Ala Leu Ile Asn Asn Gln Phe Leu Pro Tyr Pro

65 70 75 80

Tyr Tyr Ala Lys Pro Val Ala Val Arg Ser Pro Ala Gln Thr Leu Gln

85 90 95

Trp Gln Val Leu Pro Asn Thr Val Pro Ala Lys Ser Cys Gln Asp Gln

100 105 110

Pro Thr Thr Leu Ala Arg His Pro His Pro His Leu Ser Phe Met Ala

115 120 125

Ile Pro Pro Lys Lys Asp Gln Asp Lys Thr Glu Val Pro Ala Ile Asn

130 135 140

Thr Ile Ala Ser Ala Glu Pro Thr Val His Ser Thr Pro Thr Thr Glu

145 150 155 160

Ala Ile Val Asn Thr Val Asp Asn Pro Glu Ala Ser Ser Glu Ser Ile

165 170 175

Ala Ser Ala Ser Glu Thr Asn Thr Ala Gln Val Thr Ser Thr Glu Val

180 185 190

<210> 4

<211> 579

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 4

atgatgaaga gttttttcct agttgtgact atcctggcat taaccctgcc atttttgggt 60

gcccaggagc aaaaccagga acagccgata tgctgtgaga aagatgaaag attcttcgat 120

gacaaaatag ccaaatatat cccaattcag tatgtgctga gtaggtatcc tagttatgga 180

ctcaattact atcaacagag accagttgca ctaattaata atcaatttct gccataccca 240

tattatgcaa agccagttgc agttaggtca cctgcccaaa ctcttcaatg gcaagttttg 300

ccaaatactg tgcctgccaa gtcctgccaa gaccagccaa ctaccctggc acgtcaccca 360

cacccacatt tatcatttat ggccattcca ccaaagaaag atcaggataa aacagaagtc 420

cctgccatca ataccattgc tagtgctgag cctacagtac acagtacacc taccaccgaa 480

gcaatagtga acactgtaga taatccagaa gcttcctcag aatcgattgc gagtgcatct 540

gagaccaaca cagcccaagt tacttcaacc gaggtctaa 579

<210> 5

<211> 182

<212> PRT

<213> domestic cat

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 5

Met Lys Thr Phe Phe Leu Val Val Asn Ile Leu Ala Leu Thr Leu Pro

1 5 10 15

Phe Leu Gly Ala Gln Val Gln Asn Gln Glu Gln Pro Thr Cys Arg Glu

20 25 30

Asn Asp Glu Arg Leu Leu Asn Gln Lys Thr Ala Lys Tyr Ile Pro Val

35 40 45

His Tyr Val Leu Ser Asn Tyr Pro His Tyr Glu Pro Ser Tyr Tyr Pro

50 55 60

His Lys Pro Ala Val Pro Val Asn Asn Gln Tyr Met Pro Tyr Pro Tyr

65 70 75 80

Tyr Ala Lys Pro Val Ala Val Arg Pro His Val Gln Ile Pro Gln Trp

85 90 95

Gln Val Leu Pro Asn Thr Tyr Thr Pro Thr Val Val Arg His Pro His

100 105 110

Leu Pro Ala Ser Phe Ile Ala Ile Pro Pro Lys Lys Ile Gln Asp Lys

115 120 125

Thr Gly Asn Pro Thr Ile Asn Thr Ile Ala Thr Ala Glu Leu Thr Leu

130 135 140

Thr Pro Thr Thr Glu Pro Ile Val Asn Thr Val Val Thr Thr Glu Ala

145 150 155 160

Ser Ser Glu Phe Thr Ile Thr Ser Thr Pro Glu Thr Thr Thr Val Pro

165 170 175

Val Ala Ser Thr Met Val

180

<210> 6

<211> 549

<212> DNA

<213> domestic cat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 6

atgaagactt ttttcctagt tgtgaatatc cttgcattaa ccttgccatt tttgggtgca 60

caggtgcaaa accaagaaca accaacttgc cgtgaaaatg atgaaagatt gcttaatcag 120

aaaactgcca agtatatccc agttcattat gtactgagta actatcctca ctatgagccc 180

agttactacc cgcataaacc agctgtacca gttaataatc aatatatgcc ctatccatat 240

tatgcaaaac cagttgcagt taggccacat gtccaaattc ctcagtggca agtcctgcca 300

aatacctaca cacccactgt ggtacgtcac ccacacctac ctgcgtcatt tattgccatt 360

cccccaaaga aaattcagga taagacaggc aaccctacca tcaataccat tgctactgct 420

gagcttacac ttactcctac cactgaacca atagtgaaca ctgtagtcac tacagaagca 480

tcctcagaat tcaccatcac aagcacacct gagactacca cagttccagt ggcttcaacc 540

atggtctaa 549

<210> 7

<211> 196

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 7

Met Arg Ala Asn Ala Asn Val Ala Asp Ala Gln Ser Ala Ile Met Lys

1 5 10 15

Ser Phe Leu Leu Val Val Asn Ala Leu Ala Leu Thr Leu Pro Phe Leu

20 25 30

Ala Val Glu Val Gln Asn Gln Lys Gln Pro Ala Cys His Glu Asn Asp

35 40 45

Glu Arg Pro Phe Tyr Gln Lys Thr Ala Pro Tyr Val Pro Met Tyr Tyr

50 55 60

Val Pro Asn Ser Tyr Pro Tyr Tyr Gly Thr Asn Leu Tyr Gln Arg Arg

65 70 75 80

Pro Ala Ile Ala Ile Asn Asn Pro Tyr Val Pro Arg Thr Tyr Tyr Ala

85 90 95

Asn Pro Ala Val Val Arg Pro His Ala Gln Ile Pro Gln Arg Gln Tyr

100 105 110

Leu Pro Asn Ser His Pro Pro Thr Val Val Arg Arg Pro Asn Leu His

115 120 125

Pro Ser Phe Ile Ala Ile Pro Pro Lys Lys Ile Gln Asp Lys Ile Ile

130 135 140

Ile Pro Thr Ile Asn Thr Ile Ala Thr Val Glu Pro Thr Pro Ala Pro

145 150 155 160

Ala Thr Glu Pro Thr Val Asp Ser Val Val Thr Pro Glu Ala Phe Ser

165 170 175

Glu Ser Ile Ile Thr Ser Thr Pro Glu Thr Thr Thr Val Ala Val Thr

180 185 190

Pro Pro Thr Ala

195

<210> 8

<211> 591

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 8

atgagggcaa atgcaaatgt agctgatgcg caaagtgcaa taatgaagag ttttcttcta 60

gttgtcaatg ccctggcatt aaccctgcct tttttggctg tggaggttca aaaccagaaa 120

caaccagcat gccatgagaa tgatgaaaga ccattctatc agaaaacagc tccatatgtc 180

ccaatgtatt atgtgccaaa tagctatcct tattatggaa ccaatttgta ccaacgtaga 240

ccagctatag caattaataa tccatatgtg cctcgcacat attatgcaaa cccagctgta 300

gttaggccac atgcccaaat tcctcagcgg caatacctgc caaatagcca cccacccact 360

gtggtacgtc gcccaaacct gcatccatca tttattgcca tccccccaaa gaaaattcag 420

gataaaataa tcatccctac catcaatacc attgctactg ttgaacctac accagctcct 480

gccactgaac caacggtgga cagtgtagtc actccagaag ctttttcaga gtccatcatc 540

acgagcaccc ctgagacaac cacagttgca gttactccac ctacggcata a 591

<210> 9

<211> 178

<212> PRT

<213> African elephant

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 9

Met Lys Gly Phe Leu Leu Val Val Asn Ile Leu Leu Leu Pro Leu Pro

1 5 10 15

Phe Leu Ala Ala Glu Val Gln Asn Gln Glu Glu Ser Arg Cys Leu Glu

20 25 30

Lys Asp Glu Arg Trp Phe Cys Gln Lys Ala Val Lys Tyr Ile Pro Asn

35 40 45

Asp Tyr Val Leu Lys Ser Tyr Tyr Arg Tyr Glu Pro Asn Tyr Asn Gln

50 55 60

Phe Arg Ala Ala Val Pro Ile Asn Asn Pro Tyr Leu Ile Tyr Leu Tyr

65 70 75 80

Pro Ala Lys Gln Val Ala Val Arg Pro His Thr Gln Ile Pro Gln Trp

85 90 95

Gln Val Pro Ser Asn Ile Tyr Pro Ser Pro Ser Val Pro His Thr Tyr

100 105 110

Leu Lys Pro Pro Phe Ile Val Ile Pro Pro Lys Lys Thr Gln Asp Lys

115 120 125

Pro Ile Ile Pro Pro Thr Gly Thr Val Ala Ser Ile Glu Ala Thr Val

130 135 140

Glu Pro Lys Val Asn Thr Val Val Asn Ala Glu Ala Ser Ser Glu Phe

145 150 155 160

Ile Ala Thr Asn Thr Pro Glu Ala Thr Thr Val Pro Val Ile Ser Pro

165 170 175

Gln Ile

<210> 10

<211> 537

<212> DNA

<213> African elephant

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 10

atgaagggct ttctcttggt tgtgaacatc ctgttgttac ctttgccctt tttggctgca 60

gaggtgcaaa accaggagga gtcaagatgc cttgagaaag atgaaagatg gttctgtcag 120

aaagcagtca aatatattcc aaatgattat gtgctgaaga gctattatcg ttatgaacca 180

aattataacc aatttagggc agctgtacca atcaataatc catacctaat ttacctatat 240

cctgctaaac aggttgcagt taggccacat acacaaattc cgcaatggca agttccatca 300

aatatctacc catctccatc agtacctcac acatacctca aaccaccatt tattgtcatt 360

cccccaaaga aaactcagga taaacctata atccctccca ccggcaccgt tgcttctatt 420

gaagctaccg ttgagcctaa ggtgaacact gtcgtcaatg ctgaagcttc ctcagagttc 480

attgccacaa atacacctga ggctaccaca gtcccagtta tttcacccca gatctaa 537

<210> 11

<211> 175

<212> PRT

<213> mammoth

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 11

Met Lys Gly Phe Leu Leu Val Val Asn Ile Leu Leu Leu Pro Leu Phe

1 5 10 15

Leu Ala Ala Glu Val Gln Asn Gln Glu Glu Ser Arg Cys Leu Glu Lys

20 25 30

Asp Glu Arg Trp Phe Cys Gln Lys Ala Val Lys Tyr Ile Pro Asn Asp

35 40 45

Tyr Val Leu Lys Ser Tyr Tyr Arg Tyr Glu Pro Asn Tyr Asn Gln Phe

50 55 60

Arg Ala Ala Val Pro Ile Asn Asn Pro Tyr Leu Ile Tyr Leu Tyr Pro

65 70 75 80

Ala Lys Gln Val Ala Val Arg Pro His Thr Gln Ile Gln Trp Gln Val

85 90 95

Pro Ser Asn Ile Tyr Pro Ser Pro Ser Val Pro His Thr Tyr Leu Lys

100 105 110

Pro Pro Phe Ile Ile Pro Pro Lys Lys Thr Gln Asp Lys Pro Ile Ile

115 120 125

Pro Pro Thr Gly Thr Val Ala Ser Ile Glu Ala Thr Val Glu Pro Lys

130 135 140

Val Asn Thr Val Val Asn Ala Glu Ala Ser Ser Glu Phe Ile Ala Thr

145 150 155 160

Asn Thr Pro Glu Ala Thr Thr Val Pro Val Ile Ser Pro Gln Ile

165 170 175

<210> 12

<211> 546

<212> DNA

<213> mammoth

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 12

atgatgaagg gctttctctt ggttgtgaac atcctgttgt tacctttggc ctttttggct 60

gcagaggtgc aaaaccagga ggagtcaaga agctgttgcc ttgagaaaga tgaaagatgg 120

ttctgtcaga aagcagtcaa atatattcca aatgattatg tgctgaagag ctattatcgt 180

tatgaaccaa attataacca atttagggca gctgtaccaa tcaataatcc atacctaatt 240

tacctatatc ctgctaaaca ggttgcagtt aggccacata cacaaattct gcaatggcaa 300

gttccatcaa atatctaccc atctccatca gtacctcaca catacctcaa accaccattt 360

attgccattc ccccaaagaa aactcaggat aaacctataa tccctcccac cggcaccgtt 420

gcttctattg aagctaccgt tgagcctaag gtgaacactg tcgtcaatgc tgaagcttcc 480

tcagagttca ttgccacaaa tacacctgag gctaccacag tcccagttat ttcaccccag 540

atctaa 546

<210> 13

<211> 180

<212> PRT

<213> Rabbit

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 13

Met Met Lys His Phe Leu Leu Val Val Asn Ile Leu Ala Val Thr Leu

1 5 10 15

Pro Phe Leu Ala Ala Asp Ile Gln Asn Gln Glu Gln Thr Thr Cys Arg

20 25 30

Glu Asn Glu Glu Arg Leu Phe His Gln Val Thr Ala Pro Tyr Ile Pro

35 40 45

Val His Tyr Val Met Asn Arg Tyr Pro Gln Tyr Glu Pro Ser Tyr Tyr

50 55 60

Leu Arg Arg Gln Ala Val Pro Thr Leu Asn Pro Phe Met Leu Asn Pro

65 70 75 80

Tyr Tyr Val Lys Pro Ile Val Phe Lys Pro Asn Val Gln Val Pro His

85 90 95

Trp Gln Ile Leu Pro Asn Ile His Gln Pro Lys Val Gly Arg His Ser

100 105 110

His Pro Phe Phe Met Ala Ile Leu Pro Asn Lys Met Gln Asp Lys Ala

115 120 125

Val Thr Pro Thr Thr Asn Thr Ile Ala Ala Val Glu Pro Thr Pro Ile

130 135 140

Pro Thr Thr Glu Pro Val Val Ser Thr Glu Val Ile Ala Glu Ala Ser

145 150 155 160

Pro Glu Leu Ile Ile Ser Pro Glu Thr Thr Thr Glu Ala Thr Ala Ala

165 170 175

Ser Ala Ala Ala

180

<210> 14

<211> 543

<212> DNA

<213> Rabbit

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 14

atgatgaagc attttcttct agttgtgaac atcctggcag taaccttgcc ttttttggct 60

gcagacatac aaaaccagga acagacaacg tgccgtgaga atgaggaaag actgttccac 120

caggttacag ctccatatat cccagttcac tatgtgatga acagatatcc tcaatacgaa 180

cccagctact acctgcgcag acaagctgtt ccaactctta atccatttat gcttaaccca 240

tattatgtaa aaccaattgt atttaagcca aatgtccaag ttcctcactg gcaaatcctg 300

ccaaatatcc accagccaaa agtgggacgt cactcacatc cattttttat ggccattctc 360

ccgaataaaa tgcaggataa agcagtcacc cccaccacca acaccattgc tgctgtggag 420

cctaccccga ttcctaccac tgagccagtg gtgagcactg aagtgattgc agaggcttcc 480

ccagagctca tcatcagccc tgagactacc acggaagcaa ctgctgcatc agcggcagca 540

tga 543

<210> 15

<211> 192

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 15

Met Met Lys Ser Phe Phe Leu Val Val Thr Ile Leu Ala Leu Thr Leu

1 5 10 15

Pro Phe Leu Gly Ala Gln Glu Gln Asn Gln Glu Gln Arg Ile Cys Cys

20 25 30

Glu Lys Asp Glu Arg Phe Phe Asp Asp Lys Ile Ala Lys Tyr Ile Pro

35 40 45

Ile Gln Tyr Val Leu Ser Arg Tyr Pro Ser Tyr Gly Leu Asn Tyr Tyr

50 55 60

Gln Gln Arg Pro Val Ala Leu Ile Asn Asn Gln Phe Leu Pro Tyr Pro

65 70 75 80

Tyr Tyr Ala Lys Pro Val Ala Val Arg Ser Pro Ala Gln Thr Leu Gln

85 90 95

Trp Gln Val Leu Pro Asn Ala Val Pro Ala Lys Ser Cys Gln Asp Gln

100 105 110

Pro Thr Ala Met Ala Arg His Pro His Pro His Leu Ser Phe Met Ala

115 120 125

Ile Pro Pro Lys Lys Asp Gln Asp Lys Thr Glu Ile Pro Ala Ile Asn

130 135 140

Thr Ile Ala Ser Ala Glu Pro Thr Val His Ser Thr Pro Thr Thr Glu

145 150 155 160

Ala Val Val Asn Ala Val Asp Asn Pro Glu Ala Ser Ser Glu Ser Ile

165 170 175

Ala Ser Ala Pro Glu Thr Asn Thr Ala Gln Val Thr Ser Thr Glu Val

180 185 190

<210> 16

<211> 579

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 16

atgatgaaga gttttttcct agttgtgact atcctggcat taaccctgcc atttttgggt 60

gcccaggagc aaaaccaaga acaacgaata tgctgtgaga aagatgaaag attcttcgat 120

gacaaaatag ccaaatatat cccaattcag tatgtgctga gtaggtatcc tagttatgga 180

ctcaattact accaacagag accagttgca ctaattaata atcaatttct gccataccca 240

tattatgcga agccagttgc agttaggtca cctgcccaaa ctcttcaatg gcaagttttg 300

ccaaatgctg tgcctgccaa gtcctgccaa gaccagccaa ctgccatggc acgtcaccca 360

cacccacatt tatcatttat ggccattcca ccaaagaaag atcaggataa aacagaaatc 420

cctgccatca ataccattgc tagtgctgag cctacagtac acagtacacc taccaccgaa 480

gcagtagtga acgctgtaga taatccagaa gcttcctcag aatcgattgc gagtgcacct 540

gagaccaaca cagcccaagt tacttcaacc gaggtctaa 579

<210> 17

<211> 181

<212> PRT

<213> sperm whale

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 17

Met Lys Ser Phe Leu Leu Val Val Thr Ile Leu Ala Leu Thr Leu Pro

1 5 10 15

Phe Leu Ser Ala Glu Gly Gln Asn Gln Glu Gln Ser Thr Arg Cys Glu

20 25 30

Asn Asp Glu Arg Leu Phe Asn Lys Lys Thr Val Lys Tyr Ile Pro Ile

35 40 45

His Tyr Val Leu Ser Arg Tyr Pro Ser Tyr Gly Leu Asn Tyr Tyr Gln

50 55 60

His Arg Pro Val Ala Leu Ile Asn Asn Gln Phe Met Pro Tyr Leu Tyr

65 70 75 80

Tyr Ala Lys Pro Val Val Val Ser Pro His Ala Gln Ile Pro Gln Trp

85 90 95

Gln Phe Leu Pro Asn Ile His Pro Pro Thr Leu Ala His His Pro His

100 105 110

Pro Arg Pro Ser Phe Thr Ala Ile Pro Pro Lys Lys Thr Gln Asp Lys

115 120 125

Thr Ala Ile Pro Ile Ile Asn Thr Ile Ala Thr Val Glu Pro Thr Leu

130 135 140

Ile Pro Thr Thr Glu Pro Ile Val Asn Thr Val Val Thr Pro Glu Ala

145 150 155 160

Ser Ser Glu Phe Ile Thr Ser Thr Pro Glu Thr Thr Thr Val Gln Val

165 170 175

Ala Ser Pro Val Ala

180

<210> 18

<211> 546

<212> DNA

<213> sperm whale

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 18

atgaagagtt ttttactagt tgtgactatc ctggcattaa ccctgccttt tttgagtgca 60

gaggggcaaa accaggaaca atcaacacgc tgtgagaatg atgaaagatt gttcaataaa 120

aaaacagtaa aatatatccc aattcattat gtgctgagta ggtatcctag ttatggactc 180

aattactacc agcacagacc agttgcacta attaataacc aatttatgcc atacctatat 240

tatgcaaagc cagttgtagt tagcccacat gcccaaattc ctcaatggca attcctgcca 300

aatatccacc cacctactct ggcacatcac ccacacccac gtccatcatt tactgccatc 360

ccaccaaaga aaactcagga taaaacagca atccctatca tcaataccat tgctactgtt 420

gagcctacac ttatacctac cactgaacca atagtgaaca ctgtagttac tccagaagct 480

tcctcagaat tcatcacgag tacacctgag accaccacag tccaagttgc ttcacctgtg 540

gcctaa 546

<210> 19

<211> 165

<212> PRT

<213> Australia needle mole

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 19

Met Lys Thr Leu Leu Leu Val Gly Gly Ile Leu Val Met Thr Val Cys

1 5 10 15

Phe Ser Ala Ala Glu Asp Glu Glu Trp Lys Lys Val Asp Tyr Ser Glu

20 25 30

Ser Glu Glu Arg Trp Leu Arg Leu Lys Arg Gln Pro Ser Phe Pro Phe

35 40 45

Ser Phe Gln Gly Lys Pro Glu Arg Asn Ile Pro Arg Pro Tyr Tyr Pro

50 55 60

Arg Pro Phe Leu Asn Ile Pro Arg Pro Tyr Thr Ile Asn Pro Glu His

65 70 75 80

Gln Phe Ala Tyr Val Phe Pro Asn Leu Lys Phe Gln Ile Pro Ser Val

85 90 95

Phe Pro Phe Pro Leu Glu Phe Leu Pro Pro Phe Tyr Pro Phe Val His

100 105 110

Pro Ile Tyr Tyr Gly Pro Gln Thr Ser Thr Pro Pro Arg Asn Pro Thr

115 120 125

Val Thr Ser Gln Thr Pro Gln Pro Pro Val His Ser Ser Ala Asn Thr

130 135 140

Pro Glu Ser Ala Thr Ala Ala Pro Val Thr Ala Thr Pro Met Ala Gln

145 150 155 160

Thr Pro Leu Gln Pro

165

<210> 20

<211> 498

<212> DNA

<213> Australia needle mole

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 20

atgaagactc tactactggt tggaggtatc ctggttatga ccgtttgctt ctccgcggca 60

gaggacgagg aatggaaaaa ggttgattac agcgagagtg aagaaagatg gttgaggctg 120

aagcgccagc caagctttcc ctttagcttc caaggcaaac ctgagagaaa catcccacgt 180

ccttattacc ctcgaccatt tctaaatatt ccccgcccct acacgattaa ccctgagcac 240

caattcgcct atgtttttcc caacttgaag ttccaaatcc caagtgtatt tccatttccc 300

ctggaattcc tcccaccttt ctatcccttt gttcatccca tctattatgg ccctcaaacc 360

tcaacccctc ccagaaatcc caccgtgacc agccaaactc cacagccccc tgtccattcc 420

tccgccaata ccccggagtc tgctactgct gcccccgtga ctgctacccc catggcccaa 480

actcccctcc aaccttaa 498

<210> 21

<211> 178

<212> PRT

<213> broom tail bag mink

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 21

Met Lys Val Leu Phe Leu Thr Val His Ile Leu Ala Val Met Val Cys

1 5 10 15

Phe Ser Thr Ala Asp Leu Asp Trp Glu Lys Trp Pro Cys Asp Lys Gln

20 25 30

Asn Glu Arg Gln Ser Glu Leu Arg Gln Gln Pro Leu Arg Arg Ser Pro

35 40 45

Val Gln Tyr Val Tyr Thr Pro Tyr Thr His Gln Ser Tyr Val Pro Val

50 55 60

Ile Tyr Pro Pro Arg Ala Tyr Val Arg His Pro Tyr Phe Ser Arg Val

65 70 75 80

Ala Trp Gln Lys Pro Tyr Pro Ser Tyr Met Pro Leu Leu Pro Ser Ile

85 90 95

Tyr Pro Trp Ser Val Val Ser Arg Asn Leu His Pro Ala Phe Ala Phe

100 105 110

Asn Pro Pro His Tyr Ala Gln Leu Pro Val Pro Ser Ser Pro Thr Asn

115 120 125

Ser Pro Thr Thr Thr Ile Gln Thr Thr Asn Ile Pro Ile Thr Asn Pro

130 135 140

Thr Ser Thr Ile Val Thr Pro Ala Val Ser Ser Lys Ser Ala Ala Thr

145 150 155 160

Glu Asp Ser Ala Ala Ala Ala Met Leu Thr Ser Pro Thr Ala Ala Gln

165 170 175

Met Ala

<210> 22

<211> 537

<212> DNA

<213> broom tail bag mink

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 22

atgaaggtcc tattcttgac tgtgcatatt ctggcagtaa tggtgtgctt ctcaactgct 60

gacttggact gggaaaaatg gccttgcgat aagcaaaatg aaagacagtc tgagctgaga 120

caacagccac tcagacggtc ccctgtccaa tatgtctaca ccccatatac acatcaatca 180

tacgtgccag tcatttatcc accaagggca tatgtacgtc atccatattt ctctagagta 240

gcatggcaga aaccatatcc ctcctatatg ccactgctgc ccagtatcta cccttggtct 300

gtggtttcta gaaacctaca ccctgcattc gcttttaatc cccctcatta tgcccagctt 360

ccagtaccat caagtccaac caacagcccc acaactacca ttcagactac aaacattccc 420

atcactaacc ccacaagcac tatagtcacc ccagctgtct cctccaagtc tgcggccaca 480

gaggatagcg cagctgctgc aatgttgact tccccaaccg ctgctcagat ggcataa 537

<210> 23

<211> 178

<212> PRT

<213> Gray-short-tail negative mouse

<220>

<221> variants

<222> (0)...(0)

<223> Casein k

<400> 23

Met Lys Val Leu Phe Leu Ile Gly His Ile Leu Leu Ala Met Val Cys

1 5 10 15

Phe Ser Thr Ala Glu Leu Asp Trp Arg Lys Trp Pro Cys Glu Lys Gln

20 25 30

Met Glu Arg Pro Ser Glu Leu Glu Gln Gln Pro Pro Gly Gln Pro Pro

35 40 45

Val Gln Asp Val Tyr Thr Arg Tyr Thr Arg Gln Ile Tyr Val Pro Ile

50 55 60

Leu Tyr Ala Pro Lys Thr Ser Ile Gln Tyr Pro Tyr Phe Ser Lys Leu

65 70 75 80

Ala Trp Gln Arg Pro Tyr Ala Ala Tyr Ile Pro Leu Leu Ser Ser Arg

85 90 95

Tyr Pro Trp Pro Val Ile Pro Arg Ser Pro His Pro Ser Phe Ala Phe

100 105 110

Asn Pro Pro Gln Tyr Ala Arg Val Pro Ala Pro Ser Gly Pro Thr Ser

115 120 125

Ser Pro Ala Ala Pro Met Glu Thr Thr Thr Ile Pro Ser Thr Ser Thr

130 135 140

Val Ala Ala Thr Val Thr Pro Asp Ala Thr Ser Lys Phe Val Thr Thr

145 150 155 160

Glu Tyr Ser Thr Thr Ala Thr Ile Pro Thr Ser Pro Ile Pro Glu Gln

165 170 175

Gln Pro

<210> 24

<211> 537

<212> DNA

<213> Gray-short-tail negative mouse

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k cDNA

<400> 24

atgaaggtcc tgttcttgat tgggcatatt ctgttggcaa tggtgtgctt ctctactgct 60

gaactggact ggaggaaatg gccttgcgag aagcaaatgg aaagaccatc ggagctggaa 120

caacagccac ccggacagcc ccctgtccaa gacgtctaca cccgatacac ccgtcagatc 180

tacgtaccca tcttgtatgc acccaagact tccatccagt atccatattt ctctaagcta 240

gcctggcaga gaccatatgc tgcctacata ccactgctgt ccagtcgcta cccgtggcct 300

gtgattccta gaagcccaca cccttccttc gcttttaatc ccccacaata tgcccgggtt 360

ccagccccat caggtcctac cagcagcccc gcagctccca tggagactac aaccattccc 420

agcacgagca ccgtagctgc gactgtcacc cccgacgcca cttctaaatt tgtaaccacc 480

gagtatagca caactgcaac aatcccaact tccccaatcc ctgaacagca accatga 537

<210> 25

<211> 224

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 25

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Leu Glu Glu Leu Asn Val Pro Gly Glu Ile Val Glu Ser Leu Ser

20 25 30

Ser Ser Glu Glu Ser Ile Thr Arg Ile Asn Lys Lys Ile Glu Lys Phe

35 40 45

Gln Ser Glu Glu Gln Gln Gln Thr Glu Asp Glu Leu Gln Asp Lys Ile

50 55 60

His Pro Phe Ala Gln Thr Gln Ser Leu Val Tyr Pro Phe Pro Gly Pro

65 70 75 80

Ile Pro Asn Ser Leu Pro Gln Asn Ile Pro Pro Leu Thr Gln Thr Pro

85 90 95

Val Val Val Pro Pro Phe Leu Gln Pro Glu Val Met Gly Val Ser Lys

100 105 110

Val Lys Glu Ala Met Ala Pro Lys His Lys Glu Met Pro Phe Pro Lys

115 120 125

Tyr Pro Val Glu Pro Phe Thr Glu Ser Gln Ser Leu Thr Leu Thr Asp

130 135 140

Val Glu Asn Leu His Leu Pro Leu Pro Leu Leu Gln Ser Trp Met His

145 150 155 160

Gln Pro His Gln Pro Leu Pro Pro Thr Val Met Phe Pro Pro Gln Ser

165 170 175

Val Leu Ser Leu Ser Gln Ser Lys Val Leu Pro Val Pro Gln Lys Ala

180 185 190

Val Pro Tyr Pro Gln Arg Asp Met Pro Ile Gln Ala Phe Leu Leu Tyr

195 200 205

Gln Glu Pro Val Leu Gly Pro Val Arg Gly Pro Phe Pro Ile Ile Val

210 215 220

<210> 26

<211> 780

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 26

atgccattaa atactatata taaacaacca caaaatcaga tcattatcca ttcagctcct 60

ccttcacttc ttgtcctcta ctttggaaaa aaggaattga gagccatgaa ggtcctcatc 120

cttgcctgcc tggtggctct ggcccttgca agagagctgg aagaactcaa tgtacctggt 180

gagattgtgg aaagcctttc aagcagtgag gaatctatta cacgcatcaa taagaaaatt 240

gagaagtttc agagtgagga acagcagcaa acagaggatg aactccagga taaaatccac 300

ccctttgccc agacacagtc tctagtctat cccttccctg ggcccatccc taacagcctc 360

ccacaaaaca tccctcctct tactcaaacc cctgtggtgg tgccgccttt ccttcagcct 420

gaagtaatgg gagtctccaa agtgaaggag gctatggctc ctaagcacaa agaaatgccc 480

ttccctaaat atccagttga gccctttact gaaagccaga gcctgactct cactgatgtt 540

gaaaatctgc accttcctct gcctctgctc cagtcttgga tgcaccagcc tcaccagcct 600

cttcctccaa ctgtcatgtt tcctcctcag tccgtgctgt ccctttctca gtccaaagtc 660

ctgcctgttc cccagaaagc agtgccctat ccccagagag atatgcccat tcaggccttt 720

ctgctgtacc aggagcctgt actcggtcct gtccggggac ccttccctat tattgtctaa 780

<210> 27

<211> 222

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 27

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Leu Ala Ile Ala Arg

1 5 10 15

Glu Gln Glu Glu Leu Asn Val Val Gly Glu Thr Val Glu Ser Leu Ser

20 25 30

Ser Ser Glu Glu Ser Ile Thr His Ile Asn Lys Lys Ile Glu Lys Phe

35 40 45

Gln Ser Glu Glu Gln Gln Gln Thr Glu Asp Glu Leu Gln Asp Lys Ile

50 55 60

His Pro Phe Ala Gln Ala Gln Ser Leu Val Tyr Pro Phe Thr Gly Pro

65 70 75 80

Ile Pro Asn Ser Leu Pro Gln Asn Ile Leu Pro Leu Thr Gln Thr Pro

85 90 95

Val Val Val Pro Pro Phe Leu Gln Pro Glu Ile Met Gly Val Pro Lys

100 105 110

Val Lys Glu Thr Met Val Pro Lys His Lys Glu Met Pro Phe Pro Lys

115 120 125

Tyr Pro Val Glu Pro Phe Thr Glu Ser Gln Ser Leu Thr Leu Thr Asp

130 135 140

Val Glu Lys Leu His Leu Pro Leu Pro Leu Val Gln Ser Trp Met His

145 150 155 160

Gln Pro Pro Gln Pro Leu Ser Pro Thr Val Met Phe Pro Pro Gln Ser

165 170 175

Val Leu Ser Leu Ser Gln Pro Lys Val Leu Pro Val Pro Gln Lys Ala

180 185 190

Val Pro Gln Arg Asp Met Pro Ile Gln Ala Phe Leu Leu Tyr Gln Glu

195 200 205

Pro Val Leu Gly Pro Val Arg Gly Pro Phe Pro Ile Leu Val

210 215 220

<210> 28

<211> 669

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 28

atgaaggtcc tcatccttgc ctgtctggtg gctctggcca ttgcaagaga gcaggaagaa 60

ctcaatgtag tcggtgagac tgtggaaagc ctttcaagca gtgaggaatc tattacacac 120

atcaataaga aaattgagaa gtttcaaagt gaggaacaac agcaaacaga ggatgaactc 180

caggataaaa tccacccctt tgcccaggca cagtctctag tctatccctt cactgggccc 240

atccctaaca gcctcccaca aaacatcctg cctcttactc aaacccctgt ggtggtgccg 300

cctttccttc agcctgaaat aatgggagtc cccaaagtga aggagactat ggttcctaag 360

cacaaagaaa tgcccttccc taaatatcca gttgagccct ttactgaaag ccagagcctg 420

actctcactg atgttgaaaa gctgcacctt cctctgcctc tggtccagtc ttggatgcac 480

cagcctcccc agcctctttc tccaaccgtc atgtttcctc ctcagtccgt gctgtccctt 540

tctcagccca aagttctgcc tgttccccag aaagtagtgc cccagagaga tatgcccatc 600

caggcctttc tgctgtacca ggagcctgta cttggtcctg tccggggacc cttccctatt 660

cttgtctaa 669

<210> 29

<211> 244

<212> PRT

<213> domestic cat

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 29

Met Lys Val Leu Ile Leu Ala Cys Leu Leu Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Lys Glu Glu Leu Thr Val Ser Thr Glu Thr Val Glu Ser Leu Ser

20 25 30

Ser Ser Glu Glu Ser Ile Thr His Ile Asn Lys Gln Lys Leu Glu Asn

35 40 45

Leu Lys Arg Glu Glu Gln Gln Gln Arg Gln Glu Glu Arg Gln Asn Lys

50 55 60

Ile His Pro Val Phe Gln Pro Gln Pro Leu Val Tyr Pro Tyr Ala Glu

65 70 75 80

Pro Ile Pro Tyr Pro Val Leu Pro Gln Asn Ile Leu Pro Leu Ala Gln

85 90 95

Pro Ala Met Val Leu Pro Phe Leu Gln Pro Glu Ile Met Glu Ile Pro

100 105 110

Lys Val Lys Glu Thr Ile Phe Pro Arg Arg Lys Val Met Pro Ile Leu

115 120 125

Lys Ser Pro Val Val Pro Ser Leu Asp Ser Gln Ile Val Asn Leu Pro

130 135 140

Asp Leu Glu Ser Leu His Leu Pro Leu Pro Leu Pro Leu Ser Leu Leu

145 150 155 160

Gln Pro Leu Met His Gln Ile Pro Gln Pro Leu Pro Gln Thr Thr Met

165 170 175

Leu Pro Pro Gln Pro Leu Leu Ser Ile Pro Gln Pro Lys Val Met Pro

180 185 190

Phe Pro Gln Gln Ile Val Pro Tyr Leu Gln Arg Asp Met Pro Val Gln

195 200 205

Thr Leu Leu Leu Tyr Gln Asp Ala Thr Arg Glu Ala Gln Pro Val Thr

210 215 220

Ala Pro Ala Tyr Asn Pro Val Ile Val Ser Pro Asn Leu Ile Ile Pro

225 230 235 240

Leu Ser His Leu

<210> 30

<211> 735

<212> DNA

<213> domestic cat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 30

atgaaggtcc tcatcctcgc ctgcctgctg gctcttgctc ttgcaagaga gaaggaagaa 60

ctcactgtat ccactgagac tgtggaaagt ctttcaagca gtgaggaatc tattacacac 120

atcaacaagc agaaacttga gaatcttaaa cgtgaggagc agcagcagag acaggaggaa 180

cgccagaata aaatccaccc cgttttccag ccacagcctc tagtctatcc ttatgctgag 240

cccattcctt accctgttct tccacagaac atccttcccc ttgctcagcc tgctatggtg 300

ctgcctttcc ttcagcctga aataatggaa atccccaaag ttaaggagac catctttccc 360

aggcgcaaag tgatgcccat tctgaaatct ccagtagtgc cctctttgga cagccaaatc 420

gtgaatctcc ctgatcttga aagtctgcac ttgcctctgc ctctgcctct gtctctactc 480

cagcccctga tgcaccagat cccccagcct cttcctcaga ctaccatgct tcctcctcag 540

ccactgctgt ccatcccaca gcccaaagtc atgccttttc cccagcaaat tgtgccctac 600

ctccagagag acatgcccgt gcaaacccta ctgctgtacc aggatgccac ccgtgaggcc 660

caacctgtga ctgccccagc ttacaatcct gttattgtaa gtccaaactt aataattccg 720

ctgtctcact tatga 735

<210> 31

<211> 225

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 31

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Thr Ile Glu Ser Leu Ser Ser Ser Glu Glu Ser Ile Thr Glu Tyr

20 25 30

Lys Lys Val Glu Lys Val Lys His Glu Asp Gln Gln Gln Gly Glu Asp

35 40 45

Glu His Gln Asp Lys Ile Tyr Pro Ser Phe Gln Pro Gln Pro Leu Ile

50 55 60

Tyr Pro Phe Val Glu Pro Ile Pro Tyr Gly Phe Leu Pro Gln Asn Ile

65 70 75 80

Leu Pro Leu Ala Gln Pro Ala Val Val Leu Pro Val Pro Gln Pro Glu

85 90 95

Ile Met Glu Val Pro Lys Ala Lys Asp Thr Val Tyr Thr Lys Gly Arg

100 105 110

Val Met Pro Val Leu Lys Ser Pro Thr Ile Pro Phe Phe Asp Pro Gln

115 120 125

Ile Pro Lys Leu Thr Asp Leu Glu Asn Leu His Leu Pro Leu Pro Leu

130 135 140

Leu Gln Pro Leu Met Gln Gln Val Pro Gln Pro Ile Pro Gln Thr Leu

145 150 155 160

Ala Leu Pro Pro Gln Pro Leu Trp Ser Val Pro Gln Pro Lys Val Leu

165 170 175

Pro Ile Pro Gln Gln Val Val Pro Tyr Pro Gln Arg Ala Val Pro Val

180 185 190

Gln Ala Leu Leu Leu Asn Gln Glu Leu Leu Leu Asn Pro Thr His Gln

195 200 205

Ile Tyr Pro Val Thr Gln Pro Leu Ala Pro Val His Asn Pro Ile Ser

210 215 220

Val

225

<210> 32

<211> 678

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 32

atgaaggtcc tcatcctcgc ctgcctggtg gctcttgctc ttgcaaggga gaccatagaa 60

agcctttcaa gcagtgagga atctattaca gaatacaaga aagttgagaa ggttaaacat 120

gaggaccagc agcaaggaga ggatgaacac caggataaaa tctacccctc tttccagcca 180

cagcctctga tctatccatt cgttgaacct atcccctatg gttttcttcc acaaaacatt 240

ctgcctcttg ctcagcctgc tgtggtgctg cctgtccctc agcctgaaat aatggaagtc 300

cctaaagcta aagacactgt ctacactaag ggcagagtga tgcctgtcct taaatctcca 360

acgataccct tttttgaccc tcaaatccca aaactcactg atcttgaaaa tctgcatctt 420

cctctgcctc tgctccagcc cttgatgcag caggtccctc agcctattcc tcagactctt 480

gcacttcccc ctcagcccct gtggtctgtt cctcagccca aagtcctgcc tatcccccag 540

caagtggtgc cctaccctca gagagctgtg cctgttcaag cccttctgct caaccaagaa 600

cttctactta accccaccca ccagatctac cctgtgactc agccacttgc cccagttcat 660

aaccccatta gtgtctaa 678

<210> 33

<211> 263

<212> PRT

<213> African elephant

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 33

Met Lys Val Phe Ile Leu Ala Cys Leu Val Ala Phe Ala Leu Gly Arg

1 5 10 15

Glu Thr Val Glu Asn Leu Ser Ser Ser Glu Ile Arg Gln Phe Tyr Ser

20 25 30

Glu Gln Lys Pro Glu Gly Val Lys His Glu Glu Gln Gln Arg Glu Asp

35 40 45

Glu His Gln Asn Lys Ile Gln Pro Leu Phe Gln Pro Gln Pro Leu Val

50 55 60

Tyr Pro Phe Ala Glu Pro Ile Pro Tyr Thr Val Phe Pro Pro Asn Ala

65 70 75 80

Ile Pro Leu Ala Gln Pro Ile Val Val Leu Pro Phe Pro Gln Pro Glu

85 90 95

Val Lys Gln Leu Pro Glu Ala Lys Glu Ile Thr Phe Pro Arg Gln Lys

100 105 110

Leu Met Ser Phe Leu Lys Ser Pro Val Met Pro Phe Phe Asp Pro Gln

115 120 125

Ile Pro Asn Leu Gly Thr Asp Leu Glu Asn Leu His Leu Pro Leu Pro

130 135 140

Leu Leu Gln Pro Leu Arg His Gln Leu His Gln Pro Leu Ala Gln Thr

145 150 155 160

Pro Val Leu Pro Leu Pro Leu Ser Leu Pro Lys Val Leu Pro Val Pro

165 170 175

Gln Gln Val Ile Pro Tyr Pro Gln Arg Gly Arg Pro Ile Gln Asn Leu

180 185 190

Gln Leu Tyr Glu Glu Pro Leu Leu Asp Pro Thr Arg Lys Ile Tyr Pro

195 200 205

Val Ala Gln Pro Leu Ala Pro Val Tyr Asn Pro Val Ala Tyr Met Ile

210 215 220

Gly Ile Pro Cys Cys Ser Thr Leu Leu Thr Tyr Leu His Gln Ser Ser

225 230 235 240

Arg Ser Gln Tyr Pro Ile Gln Asn Lys Leu Gly Tyr Leu Ile Ala Met

245 250 255

Pro Lys Lys Val Arg Pro Thr

260

<210> 34

<211> 792

<212> DNA

<213> African elephant

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 34

atgaaggtct tcatccttgc ctgcctggtg gcttttgctc ttggaagaga gactgtagaa 60

aatctttcaa gcagtgagat aagacaattt tattcagagc aaaaacctga gggagttaag 120

catgaggaac agcaaagaga ggatgaacat cagaataaaa tccagcccct tttccagcca 180

cagcctctag tctatccttt cgctgagccc attccttata ctgtctttcc accaaacgcc 240

attcctcttg ctcagcctat tgtggtgctg cctttccctc agcctgaagt aaagcaactc 300

cctgaagcta aagaaatcac ctttcctagg caaaaattga tgtccttcct taagtctcca 360

gtaatgccct tttttgatcc ccagatccca aatcttggga ccgatcttga aaatctgcac 420

cttcctctgc ctctgctcca gcccttaaga caccagctcc accagcctct tgctcagact 480

ccagtgcttc ctcttcctct atccttgccc aaagtcctgc ccgttcccca gcaggtgata 540

ccctatcccc agagaggtag acccatccaa aaccttcaac tgtacgaaga gcctctactt 600

gacccaaccc gtaagatcta cccagtggct caaccacttg ctccagttta taaccctgtt 660

gcttacatga taggtattcc ctgttgctca acattgctca catacttaca ccagagcagt 720

aggagccagt atcccatcca gaataaactg ggttacttaa tagcaatgcc aaagaaagta 780

cgacctacat ga 792

<210> 35

<211> 302

<212> PRT

<213> Eugenia kangaroo

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 35

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Leu Gly Phe Ala Arg

1 5 10 15

Pro Met Val Glu Lys Ile Ser Glu Ser Glu Glu Tyr Val Asn Glu Val

20 25 30

Pro Glu Lys Arg Leu Lys Arg Arg Phe Pro Val Lys Asn Glu His Gln

35 40 45

Val Glu Ile Asn His His Leu Arg Pro Glu Ser Glu Met Met Ser Leu

50 55 60

Tyr Tyr Gln Pro Phe Tyr Trp Ser Glu Glu Met Arg Asn Leu Lys Met

65 70 75 80

Thr Ser Leu Pro Lys Asp Arg Arg Met Ala Val Leu Lys Ser Thr Val

85 90 95

Ser Asp Glu Val Phe Pro Ser Leu Gln His Lys Ser Leu Ser Leu Pro

100 105 110

Lys Ser Lys Val Gln Pro Leu Ser Arg Gln Gln Ile Leu Thr Phe His

115 120 125

Thr Leu Gln Met Val Pro Leu Ser His Lys Leu Leu Thr Thr Pro Lys

130 135 140

Arg Glu Met Leu Pro Ile Tyr Glu Arg Glu Arg Leu Pro Ala His Lys

145 150 155 160

Arg Glu Ser Leu Leu Ala His Glu Arg Glu Ser Leu Leu Ala His Glu

165 170 175

Arg Asp Ile Leu Val Pro Gln Arg Glu Met Ser Phe Val Pro Glu Arg

180 185 190

Glu Phe Leu Phe Ala Ser Glu Arg Val Val Leu Pro Glu Gln Glu Lys

195 200 205

Glu Ile Leu His Asn Asp Glu Arg Glu Val Leu Ala Val His Lys Lys

210 215 220

Glu Ile Leu Pro Pro Phe Glu Lys Glu Lys Val Leu Pro Leu Leu Gln

225 230 235 240

His Arg Val Val Pro Leu Pro Gln Arg Glu Ile Val Pro Pro Phe Gln

245 250 255

Arg Glu Thr Leu Leu Pro Glu Glu Ile Leu Pro Val Asn Gln Trp Glu

260 265 270

Leu Met Pro Glu Val Val Pro Phe Asp Pro Tyr Pro Phe Leu Gln Pro

275 280 285

Val Ala Pro Phe Tyr Tyr Ser Thr Glu Leu Asn Glu Lys Asn

290 295 300

<210> 36

<211> 263

<212> PRT

<213> Gray-short-tail negative mouse

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 36

Met Lys Leu Leu Ile Leu Ser Cys Leu Val Ala Leu Ala Val Ala Arg

1 5 10 15

Pro Met Val Glu Lys Ile Ser Glu Thr Glu Glu Phe Val Thr Val Ile

20 25 30

Pro Glu Gln Gln Ile Arg Arg Glu Asp Val Pro Val Lys Asn Glu Arg

35 40 45

His Pro Glu Ile Asn Arg Phe Ile Pro Leu Glu Ala Glu Thr Met Ser

50 55 60

Phe Tyr Val Pro Val Tyr Trp Pro Glu Glu Met Arg Asp Ala Lys Met

65 70 75 80

Thr Ser Pro Leu Lys Glu Lys Arg Met Thr Leu Ala Asn Pro Ile Ala

85 90 95

Pro Glu Glu Glu Leu Pro His Leu Gln His Lys Ser Leu Ser Leu Ala

100 105 110

Lys Gln Arg Phe Leu Ala Ser Leu Arg Pro Lys Ala Ala Gln Pro Phe

115 120 125

Tyr Ala Pro Arg Met Ala Pro Leu Pro His Lys Leu Phe Thr Met Pro

130 135 140

Lys Glu Gln Ala Leu Pro Ile Ala Lys Arg Asp Met Leu Ser Ala Ala

145 150 155 160

Glu Leu Val Ile Pro Ala Val His Glu Arg Val Ile Pro Ala Ile Asp

165 170 175

Lys Arg Glu Pro Leu Pro Leu Leu Ala Arg Glu Met Pro Ala Leu Pro

180 185 190

Asp Lys Glu Ile Gln Gln Leu Ala Val Pro Phe Val Arg Arg Glu Ser

195 200 205

Ala Leu Pro His Gln Arg Ala Ile Val Pro Val Ala Thr Ala Ala Ala

210 215 220

Ala Val Arg Glu Ser Leu Pro Leu Val Gln Gln Glu Val Val Pro Pro

225 230 235 240

Ile Met Pro Leu Asp Val Tyr Leu Val Arg His Pro Glu Val Ser Phe

245 250 255

Tyr Asn Pro Thr Glu Lys Tyr

260

<210> 37

<211> 792

<212> DNA

<213> Gray-short-tail negative mouse

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 37

atgaagctcc tcatcctcag ctgccttgtg gctcttgctg ttgccaggcc tatggtggaa 60

aagatctcag aaactgagga atttgtcacc gtcatcccgg agcaacagat caggagagag 120

gacgtcccag taaagaacga gcgtcatcct gaaatcaacc gctttattcc ccttgaagct 180

gaaacgatga gcttttacgt gcccgtttac tggcccgaag aaatgcgtga cgccaagatg 240

accagccctc taaaagagaa gagaatgacc ctcgctaacc ctattgcccc ggaggaagag 300

ctccctcacc tgcagcacaa atctctgtct ctcgctaagc aaagattcct ggcttctctt 360

cgccccaagg cggcgcagcc cttctatgcc ccaaggatgg cccctctccc tcacaaactg 420

tttaccatgc ccaaggagca ggcgctgcct attgccaaga gagacatgct gtccgccgcc 480

gagctcgtca tccctgcagt gcacgagaga gtcattccag ccattgacaa gagagagccc 540

ctgccgcttc ttgcgagaga gatgccagct cttcccgaca aagagattca acaactggct 600

gtgcccttcg tccgcagaga gagcgcgctc cctcaccaga gagccatcgt gcctgtcgcc 660

accgccgccg ccgccgtgag ggagagcctg cctctggtcc agcaggaagt cgtgcctccc 720

atcatgcctc tcgatgtcta cctggtccgc cacccagagg tcagcttcta caatcccacc 780

gagaagtact aa 792

<210> 38

<211> 260

<212> PRT

<213> mammoth

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 38

Met Lys Val Phe Ile Leu Ala Cys Leu Val Ala Phe Ala Leu Gly Arg

1 5 10 15

Glu Thr Val Glu Asn Leu Ser Ser Ser Glu Ile Arg Gln Phe Tyr Ser

20 25 30

Glu Gln Lys Pro Glu Gly Val Lys His Glu Glu Gln Gln Arg Glu Asp

35 40 45

Glu His Gln Asn Lys Ile Gln Pro Leu Phe Gln Pro Gln Pro Leu Val

50 55 60

Tyr Pro Phe Ala Glu Pro Ile Pro Tyr Thr Val Phe Pro Pro Asn Ala

65 70 75 80

Ile Pro Leu Ala Gln Pro Ile Val Val Leu Pro Phe Pro Gln Pro Glu

85 90 95

Val Gln Leu Pro Glu Ala Lys Glu Ile Thr Phe Pro Arg Gln Lys Leu

100 105 110

Met Ser Phe Leu Lys Ser Pro Val Met Pro Phe Phe Asp Pro Gln Pro

115 120 125

Asn Leu Gly Thr Asp Leu Glu Asn Leu His Leu Pro Leu Pro Leu Leu

130 135 140

Gln Pro Leu Arg His Gln Leu His Gln Pro Leu Ala Gln Thr Pro Val

145 150 155 160

Leu Pro Leu Pro Leu Ser Leu Pro Lys Val Leu Pro Val Pro Gln Gln

165 170 175

Val Ile Pro Tyr Pro Gln Arg Gly Arg Pro Ile Gln Asn Leu Leu Tyr

180 185 190

Glu Glu Pro Leu Leu Asp Pro Thr Arg Lys Ile Tyr Pro Val Ala Gln

195 200 205

Pro Leu Ala Pro Val Tyr Asn Pro Val Ala Tyr Met Ile Gly Ile Pro

210 215 220

Cys Cys Ser Thr Leu Leu Thr Tyr Leu His Gln Ser Ser Arg Ser Gln

225 230 235 240

Tyr Pro Ile Gln Asn Lys Leu Gly Tyr Leu Ile Ala Met Pro Lys Lys

245 250 255

Val Arg Pro Thr

260

<210> 39

<211> 717

<212> DNA

<213> mammoth

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 39

atgaaggtct tcatccttgc ctgcctggtg gcttttgctc ttggaagaga gaaggaagaa 60

attattgtat ctactgagac tgtagaaaat ctttcaagca gtgagataag gcaattttat 120

tcagaggaat ctgttacaca agtcaacaaa caaaaacctg agggagttaa gcatgaggaa 180

cagcaaagag aggatgaaca tcagaataaa atccagcccc ttttccagcc acagcctcta 240

gtctatcctt tcgctgagcc cattccttat actgtctttc caccaaacgc cattcctctt 300

gctcagccta ttgtggtgct gcctttccct cagcctgaag taatgcaact ccctgaagct 360

aaagaaatca cctttcctag gcaaaaattg atgtccttcc ttaagtctcc agtaatgccc 420

ttttttgacc cccagatgcc aaatcttggg accgatcttg aaaatctgca ccttcctctg 480

cctctactcc agcccttaag acaccagctc caccagcctc ttgctcagac tccagtgctt 540

cctcttcctc tatccttgcc caaagtcctg cccgttcccc agcaggtgat accctatccc 600

cagagaggta gacccatcca aaaccttcca ctgtacgaag agcctctact tgacccaacc 660

cgtaagatct acccagtggc tcaaccactt gctccagttt ataaccctgt tgctgta 717

<210> 40

<211> 228

<212> PRT

<213> Rabbit

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 40

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Lys Glu Gln Leu Ser Val Pro Thr Glu Ala Val Gly Ser Val Ser

20 25 30

Ser Ser Glu Glu Ile Thr His Ile Asn Lys Gln Lys Leu Glu Thr Ile

35 40 45

Lys His Val Glu Gln Leu Leu Arg Glu Glu Lys Leu Gln Asp Lys Ile

50 55 60

Leu Pro Phe Ile Gln Ser Leu Phe Pro Phe Ala Glu Arg Ile Pro Tyr

65 70 75 80

Pro Thr Leu Pro Gln Asn Ile Leu Asn Leu Ala Gln Leu Asp Met Leu

85 90 95

Leu Pro Leu Leu Gln Pro Glu Ile Met Glu Asp Pro Lys Ala Lys Glu

100 105 110

Thr Ile Ile Pro Lys His Lys Leu Met Pro Phe Leu Lys Ser Pro Lys

115 120 125

Thr Val Pro Phe Val Asp Ser Gln Ile Leu Asn Leu Arg Glu Met Lys

130 135 140

Asn Gln His Leu Leu Leu Pro Gln Leu Leu Pro Phe Met His Gln Val

145 150 155 160

Phe Gln Pro Phe Pro Gln Thr Pro Ile Pro Tyr Pro Gln Ala Leu Leu

165 170 175

Ser Leu Pro Gln Ser Lys Phe Met Pro Ile Val Pro Gln Val Val Pro

180 185 190

Tyr Pro Gln Arg Asp Met Pro Ile Gln Ala Leu Gln Leu Phe Gln Glu

195 200 205

Leu Leu Phe Pro Thr His Gln Gly Tyr Pro Val Val Gln Pro Ile Ala

210 215 220

Pro Val Asn Val

225

<210> 41

<211> 687

<212> DNA

<213> Rabbit

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 41

atgaaggtcc tcattcttgc ctgcctggtg gctctcgctc ttgcaaggga gaaggaacaa 60

ctcagtgttc ccacagaggc tgtaggaagt gtttccagca gcgaggaaat tacacatatc 120

aacaagcaga aactcgagac gattaagcac gtggaacagc tgctaagaga ggagaaactc 180

caggataaaa tcctcccctt tattcaatca ctctttcctt ttgctgagcg catcccctac 240

cctactcttc cacagaacat cctgaacctt gctcaactcg acatgctgct acctctcctt 300

cagcctgaaa taatggaaga ccccaaggct aaagagacca ttatccctaa gcacaaactg 360

atgcccttcc ttaaatctcc aaagacggtc ccctttgttg actctcaaat tctgaatctc 420

agggagatga aaaatcaaca ccttcttttg ccccagctcc tgcccttcat gcaccaggtc 480

ttccagcctt ttccccagac tcccattcca tatcctcagg ccctcctctc tcttcctcag 540

tccaaattca tgcctattgt cccacaagtg gtgccctacc ctcaaaggga catgcctatc 600

caagcccttc agctgttcca agaactgctt ttccctaccc atcaaggcta ccctgtggtt 660

caaccaatag ccccagttaa tgtctaa 687

<210> 42

<211> 222

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 42

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Gln Glu Glu Leu Asn Val Val Gly Glu Thr Val Glu Ser Leu Ser

20 25 30

Ser Ser Glu Glu Ser Ile Thr His Ile Asn Lys Lys Ile Glu Lys Phe

35 40 45

Gln Ser Glu Glu Gln Gln Gln Thr Glu Asp Glu Leu Gln Asp Lys Ile

50 55 60

His Pro Phe Ala Gln Ala Gln Ser Leu Val Tyr Pro Phe Thr Gly Pro

65 70 75 80

Ile Pro Asn Ser Leu Pro Gln Asn Ile Leu Pro Leu Thr Gln Thr Pro

85 90 95

Val Val Val Pro Pro Phe Leu Gln Pro Glu Ile Met Gly Val Pro Lys

100 105 110

Val Lys Glu Thr Met Val Pro Lys His Lys Glu Met Pro Phe Pro Lys

115 120 125

Tyr Pro Val Glu Pro Phe Thr Glu Ser Gln Ser Leu Thr Leu Thr Asp

130 135 140

Val Glu Lys Leu His Leu Pro Leu Pro Leu Val Gln Ser Trp Met His

145 150 155 160

Gln Pro Pro Gln Pro Leu Pro Pro Thr Val Met Phe Pro Pro Gln Ser

165 170 175

Val Leu Ser Leu Ser Gln Pro Lys Val Leu Pro Val Pro Gln Lys Ala

180 185 190

Val Pro Gln Arg Asp Met Pro Ile Gln Ala Phe Leu Leu Tyr Gln Glu

195 200 205

Pro Val Leu Gly Pro Val Arg Gly Pro Phe Pro Ile Leu Val

210 215 220

<210> 43

<211> 669

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<220>

<221> misc_feature

<222> 19

<223> n = A, T, C or G

<400> 43

atgaaggtcc tcatccttnc ctgtctggtg gctctggccc ttgcaagaga gcaggaagaa 60

ctcaatgtag tcggtgagac tgtggaaagc ctttcaagca gtgaggaatc tattacacac 120

atcaataaga aaattgagaa gtttcaaagt gaggaacaac agcaaacaga ggatgaactc 180

caggataaaa tccacccctt tgcccaggca cagtctctag tctatccctt cactgggccc 240

atccctaaca gcctcccaca aaacatcctg cctcttactc aaacccctgt ggtggtgccg 300

cctttccttc agcctgaaat aatgggagtc cccaaagtga aggagactat ggttcctaag 360

cacaaggaaa tgcccttccc taaatatcca gttgagccct ttactgaaag ccagagcctg 420

actctcactg atgttgaaaa gctgcacctt cctctgcctc tggtccagtc ttggatgcac 480

cagcctcccc agcctcttcc tccaaccgtc atgtttcctc ctcagtccgt gctgtccctt 540

tctcagccca aagttctgcc tgttccccag aaagcagtgc cccagagaga tatgcccatc 600

caggcctttc tgctgtacca ggagcctgta cttggtcctg tccggggacc cttccctatt 660

cttgtctaa 669

<210> 44

<211> 224

<212> PRT

<213> sperm whale

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 44

Met Lys Val Leu Ile Leu Ala Cys Leu Leu Ala Leu Ala Leu Ala Arg

1 5 10 15

Glu Lys Glu Glu Leu Asn Val Ser Gly Glu Thr Val Lys Ser Leu Ser

20 25 30

Ser Ser Glu Glu Ser Val Thr His Met Asn Lys Lys Ile Gly Lys Phe

35 40 45

Lys His Glu Glu Gln Gln Gln Thr Glu Asp Glu Arg Gln Asp Lys Ile

50 55 60

His Arg Phe Ser Gln Pro Gln Pro Leu Val Tyr Ser Tyr Thr Gly Pro

65 70 75 80

Ile Pro Tyr Pro Ile Leu Pro Gln Asn Ile Leu Pro Leu Ala Gln Pro

85 90 95

Pro Val Leu Val Pro Phe Pro Gln Pro Gly Ile Met Glu Val Pro Lys

100 105 110

Ala Lys Glu Thr Leu Leu Pro Lys His Lys Glu Met Pro Phe Pro Lys

115 120 125

Ser Pro Val Glu Pro Phe Ile Glu Ser Gln Ser Leu Thr Leu Asn Asp

130 135 140

Leu Glu Asn Leu His Leu Pro Leu Pro Leu Leu Gln Ser Leu Met His

145 150 155 160

Gln Pro Pro His Pro Leu Pro Pro Thr Pro Met Phe Pro Pro Gln Pro

165 170 175

Leu Gln Ser Leu Ser Gln Pro Lys Val Leu Pro Ile Pro Gln Gln Val

180 185 190

Val Pro Tyr Leu Gln Arg Asp Met Pro Ile Gln Ala Leu Leu Leu Tyr

195 200 205

Gln Glu Pro Val Leu Gly Pro Ile Arg Gly Leu Tyr Pro Val Ile Val

210 215 220

<210> 45

<211> 675

<212> DNA

<213> sperm whale

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 45

atgaaggtcc tcatccttgc ctgcttgttg gctcttgccc ttgcaagaga gaaagaagaa 60

ctcaatgtat ccggtgagac tgtgaaaagc ctttcaagca gtgaggaatc tgttacgcac 120

atgaacaaga aaattgggaa gtttaaacat gaggaacagc agcaaacaga ggatgaacgc 180

caggataaaa tccaccgctt ttcccagcca cagcctctag tctattccta cactgggcca 240

atcccttacc ctatccttcc acaaaacatc ctgcctcttg ctcagccccc tgtgctggtg 300

cctttccctc agcctggaat aatggaagtc cccaaagcta aggagactct ccttcctaag 360

cataaagaaa tgcccttccc taaatctcca gtagagccct ttattgaaag ccagagcctg 420

actctcaatg atcttgaaaa tctgcacctt cctctgcctc tgctccagtc cttgatgcac 480

cagcctcccc atcctcttcc tcctaccccc atgtttcctc ctcagcccct gcagtccctt 540

tctcagccca aagtcctgcc tattccccag caagtggtgc cctacctcca gagagatatg 600

cccatccagg cccttctgct gtaccaggag cctgtacttg gtcctatccg ggggctctac 660

cctgttattg tctaa 675

<210> 46

<211> 183

<212> PRT

<213> Australia needle mole

<220>

<221> variants

<222> (0)...(0)

<223> Casein beta

<400> 46

Met Lys Val Phe Ile Leu Ala Cys Leu Val Ala Val Ala Met Ala Leu

1 5 10 15

Pro Lys Gln His Ser Ser Ser Ser Ser Ser Glu Glu Ser Asp Arg Leu

20 25 30

Leu Val Lys Asp Ile Pro Thr Ala Phe Ser Ser Glu Glu His Ser Val

35 40 45

Asp Pro Lys Glu Leu Tyr Glu Pro Arg Gln Ser Tyr Ser Tyr Pro Trp

50 55 60

Gln Ser Val Arg Pro Ile Asn Thr Tyr Thr Tyr Pro Arg Ala Tyr Gln

65 70 75 80

Ile Pro Ala Val Leu Pro Met Thr His Pro Gln Thr Leu Thr Tyr Leu

85 90 95

Gln Pro Gln Phe Lys Pro Glu Asp Met Ser Ile Ser Gln Lys Gln Ile

100 105 110

Pro Pro Tyr Val Gln Ala Val Val Met Pro Tyr Pro Gln Val Glu Ala

115 120 125

Ile Pro Phe Pro Gly Ala Glu Phe Met Pro Tyr Ala Gln Pro Ile Thr

130 135 140

Thr Pro Leu Leu Gln Pro Glu Val Phe Ser Ala Pro Phe Tyr Arg Glu

145 150 155 160

Ala Val Phe Lys Pro Val Ile Tyr Gly Leu Pro Gln Ser Gln Pro Val

165 170 175

Gln Lys Ile Pro Glu Thr Asp

180

<210> 47

<211> 552

<212> DNA

<213> Australia needle mole

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta cDNA

<400> 47

atgaaggtct tcatcctcgc ctgcctagtg gctgttgcca tggcattgcc taaacaacac 60

agcagcagct cttccagtga ggaatctgac agactgctgg ttaaggacat tcctactgcc 120

ttctccagcg aggaacactc tgtggacccg aaggaactct acgagccccg tcagagctat 180

tcctacccat ggcaatcagt ccgtcccatc aacacctaca cttatcctcg cgcttaccaa 240

attccggctg tcctccccat gactcatcct cagaccctga cttatctcca gcctcaattc 300

aagcccgaag acatgtctat ttctcagaaa caaatcccgc cctacgtcca ggctgtagtc 360

atgccctacc cccaggttga agccattcct ttccccgggg ctgaattcat gccctacgct 420

caacccatca ccacgcctct acttcagcct gaggtcttct ccgccccatt ctacagagag 480

gccgtcttca agccagtgat ctacggcctt cctcaatctc aaccagttca gaagatccca 540

gaaaccgact ga 552

<210> 48

<211> 214

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 48

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys His Pro Ile Lys His Gln Gly Leu Pro Gln Glu Val Leu Asn

20 25 30

Glu Asn Leu Leu Arg Phe Phe Val Ala Pro Phe Pro Glu Val Phe Gly

35 40 45

Lys Glu Lys Val Asn Glu Leu Ser Lys Asp Ile Gly Ser Glu Ser Thr

50 55 60

Glu Asp Gln Ala Met Glu Asp Ile Lys Gln Met Glu Ala Glu Ser Ile

65 70 75 80

Ser Ser Ser Glu Glu Ile Val Pro Asn Ser Val Glu Gln Lys His Ile

85 90 95

Gln Lys Glu Asp Val Pro Ser Glu Arg Tyr Leu Gly Tyr Leu Glu Gln

100 105 110

Leu Leu Arg Leu Lys Lys Tyr Lys Val Pro Gln Leu Glu Ile Val Pro

115 120 125

Asn Ser Ala Glu Glu Arg Leu His Ser Met Lys Glu Gly Ile His Ala

130 135 140

Gln Gln Lys Glu Pro Met Ile Gly Val Asn Gln Glu Leu Ala Tyr Phe

145 150 155 160

Tyr Pro Glu Leu Phe Arg Gln Phe Tyr Gln Leu Asp Ala Tyr Pro Ser

165 170 175

Gly Ala Trp Tyr Tyr Val Pro Leu Gly Thr Gln Tyr Thr Asp Ala Pro

180 185 190

Ser Phe Ser Asp Ile Pro Asn Pro Ile Gly Ser Glu Asn Ser Gly Lys

195 200 205

Thr Thr Met Pro Leu Trp

210

<210> 49

<211> 645

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 49

atgaaacttc tcatccttac ctgtcttgtg gctgttgctc ttgctaggcc taaacatcct 60

atcaagcacc aaggactccc tcaagaagtc ctcaatgaaa atttactcag gttttttgtg 120

gcaccttttc cagaagtgtt tggaaaggag aaggtcaatg aactgagcaa ggatattggg 180

agtgaatcaa ctgaggatca agccatggaa gatattaagc aaatggaagc tgaaagcatt 240

tcgtcaagtg aggaaattgt tcccaatagt gttgagcaga agcacattca aaaggaagat 300

gtgccctctg agcgttacct gggttatctg gaacagcttc tcagactgaa aaaatacaaa 360

gtaccccagc tggaaattgt tcccaatagt gctgaggaac gacttcacag tatgaaagag 420

ggaatccatg cccaacagaa agaacctatg ataggagtga atcaggaact ggcctacttc 480

taccctgagc ttttcagaca attctaccag ctggatgcct atccatctgg tgcctggtat 540

tacgttccac taggcacaca atacactgat gccccatcat tctctgacat ccctaatcct 600

attggctctg agaacagtga aaagactact atgccactgt ggtga 645

<210> 50

<211> 222

<212> PRT

<213> Camel with Single Peak

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 50

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys Tyr Pro Leu Arg Tyr Pro Glu Val Phe Gln Asn Glu Pro Asp

20 25 30

Ser Ile Glu Glu Val Leu Asn Lys Arg Lys Ile Leu Asp Leu Ala Val

35 40 45

Val Ser Pro Ile Gln Phe Arg Gln Glu Asn Ile Asp Glu Leu Lys Asp

50 55 60

Thr Arg Asn Glu Pro Thr Glu Asp His Ile Met Glu Asp Thr Glu Arg

65 70 75 80

Lys Glu Ser Gly Ser Ser Ser Ser Glu Glu Val Val Ser Ser Thr Thr

85 90 95

Glu Gln Lys Asp Ile Leu Lys Glu Asp Met Pro Ser Gln Arg Tyr Leu

100 105 110

Glu Glu Leu His Arg Leu Asn Lys Tyr Lys Leu Leu Gln Leu Glu Ala

115 120 125

Ile Arg Asp Gln Lys Leu Ile Pro Arg Val Lys Leu Ser Ser His Pro

130 135 140

Tyr Leu Glu Gln Leu Tyr Arg Ile Asn Glu Asp Asn His Pro Gln Leu

145 150 155 160

Gly Glu Pro Val Lys Val Val Thr Gln Pro Phe Pro Gln Phe Phe Gln

165 170 175

Leu Gly Ala Ser Pro Tyr Val Ala Trp Tyr Tyr Pro Pro Gln Val Met

180 185 190

Gln Tyr Ile Ala His Pro Ser Ser Tyr Asp Thr Pro Glu Gly Ile Ala

195 200 205

Ser Glu Asp Gly Gly Lys Thr Asp Val Met Pro Gln Trp Trp

210 215 220

<210> 51

<211> 669

<212> DNA

<213> Camel with Single Peak

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 51

atgaagcttc tcatccttac ctgccttgtg gctgttgcgc ttgccaggcc taaatatcct 60

ctcaggtacc cagaagtctt tcaaaatgaa ccagacagca tagaggaagt cctcaacaaa 120

agaaagattc ttgatttagc agtggtttca cccattcagt ttagacagga gaacatcgat 180

gaactgaagg atactaggaa cgaaccaacc gaagatcaca tcatggaaga cactgagcga 240

aaggaatctg gaagcagttc aagtgaggaa gttgtttcca gtaccactga gcagaaggac 300

attctcaagg aagatatgcc ctcccaacgc tatctggaag agcttcacag actgaacaaa 360

tacaaactac tccagctgga agctatccgt gaccagaaac ttattccaag agtgaagctg 420

tcctcccacc catatctgga acaactttac agaataaatg aggacaacca cccccaactg 480

ggggagcctg tgaaagtagt gactcagcct ttcccacaat tcttccagct tggtgcctct 540

ccctatgttg cttggtatta tcctccacaa gtcatgcaat atattgctca cccctcatcc 600

tacgacaccc ctgaaggcat tgcctctgag gacggtggaa aaaccgacgt tatgccacag 660

tggtggtga 669

<210> 52

<211> 213

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 52

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Val Val Ala Leu Ala Arg

1 5 10 15

Pro Lys His Pro Ile Asn His Gln Gly Leu Ser Pro Glu Val Leu Asn

20 25 30

Glu Asn Leu Leu Arg Phe Val Val Ala Pro Phe Pro Glu Val Phe Arg

35 40 45

Lys Glu Asn Ile Asn Glu Leu Ser Lys Asp Ile Gly Ser Glu Ser Thr

50 55 60

Glu Asp Gln Ala Met Glu Asp Ala Lys Gln Met Lys Ala Gly Ser Ser

65 70 75 80

Ser Ser Ser Glu Glu Ile Val Pro Asn Ser Ala Gln Lys Tyr Ile Gln

85 90 95

Lys Glu Asp Val Pro Ser Glu Arg Tyr Leu Gly Tyr Leu Glu Gln Leu

100 105 110

Leu Arg Leu Lys Lys Tyr Asn Val Pro Gln Leu Glu Ile Val Pro Lys

115 120 125

Ser Ala Glu Glu Gln Leu His Ser Met Lys Glu Gly Asn Pro Ala His

130 135 140

Gln Lys Gln Pro Met Ile Ala Val Asn Gln Glu Leu Ala Tyr Phe Tyr

145 150 155 160

Pro Gln Leu Phe Arg Gln Phe Tyr Gln Leu Asp Ala Tyr Pro Ser Gly

165 170 175

Ala Trp Tyr Tyr Leu Pro Leu Gly Thr Gln Tyr Thr Asp Ala Pro Ser

180 185 190

Phe Ser Asp Ile Pro Asn Pro Ile Gly Ser Glu Asn Ser Gly Lys Thr

195 200 205

Thr Met Pro Leu Trp

210

<210> 53

<211> 642

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 53

atgaaacttc tcatccttac ctgtcttgtg gttgttgctc ttgccaggcc taaacatcct 60

atcaatcacc aaggactctc tccagaagtc ctcaatgaaa atttactcag gtttgttgtg 120

gcgccttttc cagaagtgtt tagaaaggag aacatcaatg aactgagtaa ggatattggg 180

agtgaatcaa ctgaggatca agccatggaa gatgctaagc aaatgaaagc tggaagcagt 240

tcgtcaagtg aggaaattgt tcccaatagt gctcagaagt acattcaaaa ggaagatgtg 300

ccctctgagc gttacctggg ttatctggaa cagcttctca gactgaaaaa atacaacgtg 360

ccccagctgg aaattgttcc caaaagtgct gaggaacaac ttcacagtat gaaagaggga 420

aaccctgccc accagaaaca gcctatgata gcagtgaacc aggaactggc ctacttctac 480

cctcagcttt tcagacaatt ctaccagctg gacgcctatc catctggtgc ctggtactac 540

cttccactag gcacacaata cactgatgcc ccctcattct ctgacatccc taatcccatt 600

ggctctgaga acagtggaaa gactactatg ccactgtggt ga 642

<210> 54

<211> 212

<212> PRT

<213> horse of Przewalskii

<220>

<221> CONFLICT

<222> (0)...(0)

<223> Casein alpha-S1

<400> 54

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys Leu Pro His Arg Gln Pro Glu Ile Ile Gln Asn Lys Gln Asp

20 25 30

Ser Arg Glu Lys Val Leu Lys Glu Arg Lys Phe Pro Ser Phe Ala Leu

35 40 45

Glu Tyr Ile Asn Glu Leu Asn Arg Gln Arg Glu Leu Leu Lys Glu Lys

50 55 60

Gln Lys Asp Glu His Lys Glu Tyr Leu Ile Glu Asp Pro Glu Gln Gln

65 70 75 80

Glu Ser Ser Ser Thr Ser Ser Ser Glu Glu Val Val Pro Ile Asn Thr

85 90 95

Glu Gln Lys Arg Ile Pro Arg Glu Asp Met Leu Tyr Gln His Thr Leu

100 105 110

Glu Gln Leu Arg Arg Leu Ser Lys Tyr Asn Gln Leu Gln Leu Gln Ala

115 120 125

Ile His Ala Gln Glu Gln Leu Leu Arg Met Lys Glu Asn Ser Gln Arg

130 135 140

Lys Pro Met Arg Val Val Asn Gln Glu Gln Ala Tyr Phe Tyr Leu Glu

145 150 155 160

Pro Phe Gln Pro Ser Tyr Gln Leu Asp Val Tyr Pro Tyr Ala Ala Trp

165 170 175

Phe His Pro Ala Gln Ile Met Gln His Val Ala Tyr Ser Pro Phe His

180 185 190

Asp Thr Ala Lys Leu Ile Ala Ser Glu Asn Ser Glu Lys Thr Asp Ile

195 200 205

Ile Pro Glu Trp

210

<210> 55

<211> 639

<212> DNA

<213> horse of Przewalskii

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 55

atgaagcttc tcatccttac ctgccttgtg gctgttgctc ttgccaggcc taaacttcct 60

catagacagc cagaaatcat tcagaataaa caggacagta gagagaaagt cctcaaagaa 120

agaaagtttc ccagttttgc tctagagtac atcaatgaac tgaacaggca gagagaactt 180

ctgaaagaaa aacagaaaga tgaacacaag gaatatctca tagaagaccc tgagcaacag 240

gaatctagca gcacttcatc aagcgaggaa gttgttccca ttaacactga gcagaagcgc 300

attccaaggg aagacatgct ctaccaacac actctggaac agcttcgcag actgagcaaa 360

tacaaccaac tccagctgca agccatccat gcccaggaac aacttctcag aatgaaggaa 420

aacagccaga gaaagcctat gagagtagtg aatcaggaac aggcctactt ctaccttgag 480

cctttccaac catcctacca gcttgatgtc tatccctatg ctgcttggtt tcatcctgcg 540

caaatcatgc aacatgttgc ttactcacca ttccatgaca ctgccaaact cattgcctct 600

gagaactcgg aaaaaactga cattatacca gagtggtga 639

<210> 56

<211> 212

<212> PRT

<213> Biya wild donkey

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 56

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys Leu Pro His Arg His Pro Glu Ile Ile Gln Asn Glu Gln Asp

20 25 30

Ser Arg Glu Lys Val Leu Lys Glu Arg Lys Phe Pro Ser Phe Ala Leu

35 40 45

Glu Tyr Ile Asn Glu Leu Asn Arg Gln Arg Glu Leu Leu Lys Glu Lys

50 55 60

Gln Lys Asp Glu His Lys Glu Tyr Leu Ile Glu Asp Pro Glu Gln Gln

65 70 75 80

Glu Ser Ser Ser Thr Ser Ser Ser Glu Glu Val Val Pro Ile Asn Thr

85 90 95

Glu Gln Lys Arg Ile Pro Arg Glu Asp Met Leu Tyr Gln His Thr Leu

100 105 110

Glu Ala Leu Arg Arg Leu Ser Lys Tyr Asn Gln Leu Gln Leu Gln Ala

115 120 125

Ile Tyr Ala Gln Glu Gln Leu Leu Arg Met Lys Glu Asn Ser Gln Arg

130 135 140

Lys Pro Met Arg Val Val Asn Gln Glu Gln Ala Tyr Phe Tyr Leu Glu

145 150 155 160

Pro Phe Gln Pro Ser Tyr Gln Leu Asp Val Tyr Pro Tyr Ala Ala Trp

165 170 175

Phe His Pro Ala Gln Ile Met Gln His Val Ala Tyr Ser Pro Phe His

180 185 190

Asp Thr Ala Lys Leu Ile Ala Ser Glu Asn Ser Glu Lys Thr Asp Ile

195 200 205

Ile Pro Glu Trp

210

<210> 57

<211> 212

<212> PRT

<213> horse

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 57

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys Leu Pro His Arg Gln Pro Glu Ile Ile Gln Asn Glu Gln Asp

20 25 30

Ser Arg Glu Lys Val Leu Lys Glu Arg Lys Phe Pro Ser Phe Ala Leu

35 40 45

Glu Tyr Ile Asn Glu Leu Asn Arg Gln Arg Glu Leu Leu Lys Glu Lys

50 55 60

Gln Lys Asp Glu His Lys Glu Tyr Leu Ile Glu Asp Pro Glu Gln Gln

65 70 75 80

Glu Ser Ser Ser Thr Ser Ser Ser Glu Glu Val Val Pro Ile Asn Thr

85 90 95

Glu Gln Lys Arg Ile Pro Arg Glu Asp Met Leu Tyr Gln His Thr Leu

100 105 110

Glu Gln Leu Arg Arg Leu Ser Lys Tyr Asn Gln Leu Gln Leu Gln Ala

115 120 125

Ile His Ala Gln Glu Gln Leu Ile Arg Met Lys Glu Asn Ser Gln Arg

130 135 140

Lys Pro Met Arg Val Val Asn Gln Glu Gln Ala Tyr Phe Tyr Leu Glu

145 150 155 160

Pro Phe Gln Pro Ser Tyr Gln Leu Asp Val Tyr Pro Tyr Ala Ala Trp

165 170 175

Phe His Pro Ala Gln Ile Met Gln His Val Ala Tyr Ser Pro Phe His

180 185 190

Asp Thr Ala Lys Leu Ile Ala Ser Glu Asn Ser Glu Lys Thr Asp Ile

195 200 205

Ile Pro Glu Trp

210

<210> 58

<211> 640

<212> DNA

<213> horse

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 58

catgaagctt ctcatcctta cctgccttgt ggctgttgct cttgccaggc ctaaacttcc 60

tcatagacag ccagaaatca ttcagaatga acaggacagt agagagaaag tcctcaaaga 120

aagaaagttt cccagttttg ctctagagta catcaatgaa ctgaacaggc agagagaact 180

tctgaaagaa aaacagaaag atgaacacaa ggaatatctc atagaagacc ctgagcaaca 240

ggaatctagc agcacttcat caagcgagga agttgttccc attaacactg agcagaagcg 300

cattccaagg gaagacatgc tctaccaaca cactctggaa cagcttcgca gactgagcaa 360

atacaaccaa ctccagctgc aagccatcca tgcccaggaa caacttatca gaatgaagga 420

aaacagccag agaaagccta tgagagtagt gaatcaggaa caggcctact tctaccttga 480

gcctttccaa ccatcctacc agcttgatgt ctatccctat gctgcttggt ttcatcctgc 540

gcaaatcatg caacatgttg cttactcacc attccatgac actgccaaac tcattgcctc 600

tgagaactcg gaaaaaactg acattatacc agagtggtga 640

<210> 59

<211> 125

<212> PRT

<213> domestic cat

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 59

Met Glu Asp Pro Glu Gln Arg Glu Ile Ser Ser Ser Ser Ser Ser Glu

1 5 10 15

Glu Ala Val Pro Asn Asn Thr Gln Gln Lys His Ile Ser Lys Glu Asp

20 25 30

Ile Leu Ser Gln Arg Tyr Leu Glu Gln Leu His Arg Leu Ser Lys Tyr

35 40 45

Asn Gln Leu Gln Leu Glu Ala Leu Arg Asp Gln Gln Gln Leu Arg Arg

50 55 60

Val Thr Glu Asn Asn His Ile Gln Leu Pro Phe Gln Gln Phe Tyr Gln

65 70 75 80

Leu Asp Ala Tyr Pro Tyr Ala Val Trp Tyr Tyr Pro Pro Gln Val Met

85 90 95

Gln Tyr Ile Ala Tyr Thr Pro Phe Tyr Asp Val Thr Lys Leu Thr Ala

100 105 110

Pro Glu Asn Ala Glu Asn Val Gly Val Val Pro Glu Trp

115 120 125

<210> 60

<211> 378

<212> DNA

<213> domestic cat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 60

atggaagacc ctgagcaaag ggaaattagc agcagttcat caagcgagga agctgttccc 60

aataacactc agcagaagca catttcaaag gaagatatac tctctcaacg ctatctggaa 120

cagcttcata gactgagcaa atacaaccaa ctgcaactgg aagctctccg tgaccagcaa 180

caactgcgca gagtgactga aaacaaccac atccaattgc ctttccaaca attctaccaa 240

cttgatgctt atccctatgc tgtgtggtat taccctccac aagtcatgca gtatattgct 300

tacacaccat tctatgacgt cactaaactt acggcccctg agaacgctga aaacgttggt 360

gttgtgccag agtggtag 378

<210> 61

<211> 185

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 61

Met Arg Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys Leu Pro Leu Arg Tyr Pro Glu Arg Leu Gln Asn Pro Ser Glu

20 25 30

Ser Ser Glu Pro Ile Pro Leu Glu Ser Arg Glu Glu Tyr Met Asn Gly

35 40 45

Met Asn Arg Gln Arg Asn Ile Leu Arg Glu Lys Gln Thr Asp Glu Ile

50 55 60

Lys Asp Thr Arg Asn Glu Ser Thr Gln Asn Cys Val Val Ala Glu Pro

65 70 75 80

Glu Lys Met Glu Ser Ser Ile Ser Ser Ser Ser Glu Glu Met Ser Leu

85 90 95

Ser Lys Cys Ala Glu Gln Phe Cys Arg Leu Asn Glu Tyr Asn Gln Leu

100 105 110

Gln Leu Gln Ala Val His Ala Gln Glu Gln Ile Arg Arg Met Asn Glu

115 120 125

Asn Ser His Val Gln Val Pro Phe Gln Gln Leu Asn Gln Leu Ala Ala

130 135 140

Tyr Pro Tyr Ala Val Trp Tyr Tyr Pro Gln Ile Met Gln Tyr Val Pro

145 150 155 160

Phe Pro Pro Phe Ser Asp Ile Ser Asn Pro Thr Ala His Glu Asn Tyr

165 170 175

Glu Lys Asn Asn Val Met Leu Gln Trp

180 185

<210> 62

<211> 558

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 62

atgaggcttc tcattctcac ctgtcttgtg gctgttgctc ttgccaggcc taaacttcct 60

cttagatacc cagaacgcct tcagaatcca tcagagagca gtgagcctat accattagaa 120

tcaagagagg aatacatgaa tggtatgaac aggcagagaa acattctgag agaaaaacag 180

actgatgaaa tcaaggatac taggaatgag tctactcaga actgtgttgt ggcagagcct 240

gagaagatgg aatccagcat cagttcatcg agtgaggaaa tgtctctcag taagtgtgcg 300

gaacagtttt gtagactgaa cgaatacaac caacttcagc tgcaagctgt ccatgcccag 360

gagcaaattc gcagaatgaa tgaaaacagc catgtccaag tgcctttcca gcagctcaac 420

caacttgctg cctaccccta tgctgtttgg tactatccac aaatcatgca gtatgttcct 480

ttcccaccgt tttccgacat ctccaatccc actgctcatg aaaattatga aaaaaataac 540

gtcatgctac agtggtga 558

<210> 63

<211> 237

<212> PRT

<213> Eugenia kangaroo

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 63

Met Lys Leu Leu Ile Phe Ser Cys Leu Val Thr Leu Ala Leu Ala Arg

1 5 10 15

Pro Asp Ala Leu Arg Leu Ser Ile Asp Arg His Phe Lys His Arg Glu

20 25 30

Leu Glu Asn Arg Leu Asn Glu Asp Pro Ile Pro Val Ser Glu Ala Ser

35 40 45

Ser Ser Glu Glu Ser Val His Gln Leu Asn Arg Asp Arg Arg Pro Leu

50 55 60

Glu Lys Tyr Glu Leu Asp Lys Tyr Arg Glu Asp Leu Lys Thr Ser Ser

65 70 75 80

Ser Glu Glu Phe Val Thr Pro Ser Thr Asn Glu Arg Val Arg Arg Gln

85 90 95

Val Glu Tyr Asn Phe Asn Glu Glu Asp Ser Ser Ala Ser Arg Glu Arg

100 105 110

Lys Ile Glu Asp Phe Ser Glu His Asp Arg Gln Tyr Leu Arg Arg Arg

115 120 125

Val Glu Glu Arg Ala Leu Asn Leu Arg Tyr Leu Glu Pro Leu Tyr Tyr

130 135 140

Ala Thr Glu Pro Glu Tyr Tyr Tyr Tyr Tyr Ala Tyr Val Pro Val Ser

145 150 155 160

Ser His Asp Ile Pro Tyr Gln Gln Lys Pro Leu Ser Leu Leu Pro Ala

165 170 175

Lys Ser His Tyr Leu Ile Ser Thr Gly Leu Leu Asn Glu Pro Leu Pro

180 185 190

Ile Leu Arg Glu Arg Leu Gly Arg Gly Phe Gln Ser Pro Ser Leu Leu

195 200 205

Ile Leu Val Leu Thr Glu Asn Ser Asn Leu Phe Met Gly Ser Val Phe

210 215 220

Tyr Trp Cys Leu Gln Ile Ala His Pro Met Gln Glu Ile

225 230 235

<210> 64

<211> 189

<212> PRT

<213> Gray-short-tail negative mouse

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 64

Met Lys Leu Leu Ile Phe Ser Cys Leu Val Ala Leu Ala Leu Ala Arg

1 5 10 15

Pro Glu Ala Leu Asn Phe Ser Ala Arg Arg Val Lys His Gln Glu Ala

20 25 30

Glu Ser Arg Leu Asn Glu Val Ile Ile Pro Ala Arg Ala Val Ser Ser

35 40 45

Ser Glu Glu Thr Ser Gln Glu Ala Ile Glu Ile Arg Tyr Pro Leu Glu

50 55 60

Gln Gln Val Leu Asp Lys Ala Arg Glu Glu Arg Val Arg Arg Pro Val

65 70 75 80

Glu Tyr Ile Ile Glu Asp Asp Ser Ser Ala Leu Asn Glu Arg Lys Ile

85 90 95

Glu Asp Ala Arg Ala Tyr Asp Glu Gln Tyr Leu Arg Arg Pro Glu Glu

100 105 110

Glu Arg Ala Val His Tyr Arg Glu Leu Arg Ala Phe Pro Thr Glu Ala

115 120 125

Arg Lys Leu Lys Ala Tyr Arg Glu Pro Tyr Val Gln Pro Glu Ile Tyr

130 135 140

Tyr Tyr Leu Ile Ser Val Pro Gln Pro Met Pro Tyr Pro Asp Glu Val

145 150 155 160

Pro Leu Ala Tyr Thr Tyr Lys Phe Val Val Pro Ala Val Asn Arg Ala

165 170 175

Asp Glu Ala Val Asn Thr Pro Val Glu Glu Glu Lys Asn

180 185

<210> 65

<211> 570

<212> DNA

<213> Gray-short-tail negative mouse

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 65

atgaagctgc tcatcttctc ctgccttgtg gctcttgctc tggccaggcc agaagccctc 60

aacttctctg ctaggcgtgt taaacaccaa gaagcagaaa gccgcctgaa tgaagttatc 120

attccagcaa gagcggtttc atcaagtgag gaaacttccc aggaggcaat cgaaatcaga 180

tatcccctgg agcagcaagt actcgacaaa gccagagagg aacgtgtccg cagaccggtc 240

gagtacatca tcgaggatga ttcttctgcc ttaaatgaga gaaagattga agatgcccgt 300

gcatacgacg agcagtacct gagaagacct gaggaagaga gagctgttca ctaccgggaa 360

cttcgcgctt ttcctactga ggcaagaaaa cttaaggctt acagagaacc ctacgtgcag 420

ccagaaatct actactatct catttccgtg ccacaaccca tgccttatcc agatgaggtc 480

cctcttgctt acacctacaa attcgtagta cctgctgtca acagggcaga tgaggcagtc 540

aatacccctg tggaagagga gaagaactaa 570

<210> 66

<211> 207

<212> PRT

<213> mouse lemur

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 66

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys His Pro Leu Arg His Pro Glu Leu Ile Gln Asn Gln Pro Gly

20 25 30

Ser Ser Glu Glu Ile Leu Lys Glu Arg Lys Phe Ser Ala Ile Ala Leu

35 40 45

Ala Thr Pro Ile Glu Leu Arg Gln Glu Tyr Ile Asn Glu Leu Asn Arg

50 55 60

Glu Gln His Val Ile Thr Glu Thr Glu Gln Ser Glu Ser Ser Ser Ser

65 70 75 80

Ser Ser Ser Glu Glu Val Ala Ser Gln Ser Ser Thr Glu Pro Lys Cys

85 90 95

Ala Leu Asn Glu Asp Val Thr Asn Gln Cys Asn Gln Glu Gln Leu His

100 105 110

Arg Met Asn Lys Tyr Asn Gln Leu Gln Leu Glu Ala Ile His Ala Gln

115 120 125

Glu Gln Leu Arg Arg Met Asn Glu Tyr Asn His Ala Gln Val Glu Glu

130 135 140

Pro Val Arg Val Val Asn Gln Glu Gln Ala Gln Phe Tyr Pro Glu Pro

145 150 155 160

Phe Pro Gln Val Tyr Gln Leu Asp Ala Thr Trp Tyr Tyr Phe Pro Gln

165 170 175

Asn Met Gln Tyr Pro Ser Phe Leu Pro Ser Gln Asp Ile Ala Lys Gln

180 185 190

Thr Ser Ala Glu Asn Asn Glu Lys Thr Asn Val Met Ala Gln Trp

195 200 205

<210> 67

<211> 624

<212> DNA

<213> mouse lemur

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 67

atgaagcttc tcatactcac ctgccttgtg gctgtcgctc ttgccagacc taaacatcct 60

cttagacacc cagaactcat tcaaaatcaa ccaggcagca gcgaggaaat cctcaaagaa 120

agaaagtttt cagcaattgc tctggccaca ccaatagaat taagacagga atacatcaat 180

gaactgaaca gggaacaaca tgtaataacc gagactgagc agagtgaatc tagcagcagt 240

tcatcaagcg aggaagttgc ttcccaaagc agcaccgagc caaaatgcgc tttaaatgaa 300

gatgtgacca accaatgcaa tcaggaacag cttcatagaa tgaacaaata caaccaactc 360

cagctggaag ctatccatgc tcaggagcaa cttcgcagaa tgaatgaata caaccatgct 420

caagtggaag agcccgtaag agtagtgaat caggaacagg cccaattcta ccctgagccc 480

ttccctcaag tctaccagct tgacgctact tggtactatt tcccacaaaa catgcagtac 540

ccatctttcc tgccatccca agacatcgct aagcaaactt ccgctgagaa caatgagaaa 600

actaatgtta tggcacagtg gtga 624

<210> 68

<211> 215

<212> PRT

<213> Rabbit

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 68

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Thr Ala Leu Ala Arg

1 5 10 15

His Lys Phe His Leu Gly His Leu Lys Leu Thr Gln Glu Gln Pro Glu

20 25 30

Ser Ser Glu Gln Glu Ile Leu Lys Glu Arg Lys Leu Leu Arg Phe Val

35 40 45

Gln Thr Val Pro Leu Glu Leu Arg Glu Glu Tyr Val Asn Glu Leu Asn

50 55 60

Arg Gln Arg Glu Leu Leu Arg Glu Lys Glu Asn Glu Glu Ile Lys Gly

65 70 75 80

Thr Arg Asn Glu Val Thr Glu Glu His Val Leu Ala Asp Arg Glu Thr

85 90 95

Glu Ala Ser Ile Ser Ser Ser Ser Glu Glu Ile Val Pro Ser Ser Thr

100 105 110

Lys Gln Lys Tyr Val Pro Arg Glu Asp Leu Ala Tyr Gln Pro Tyr Val

115 120 125

Gln Gln Gln Leu Leu Arg Met Lys Glu Arg Tyr Gln Ile Gln Glu Arg

130 135 140

Glu Pro Met Arg Val Val Asn Gln Glu Leu Ala Gln Leu Tyr Leu Gln

145 150 155 160

Pro Phe Glu Gln Pro Tyr Gln Leu Asp Ala Tyr Leu Pro Ala Pro Trp

165 170 175

Tyr Tyr Thr Pro Glu Val Met Gln Tyr Val Leu Ser Pro Leu Phe Tyr

180 185 190

Asp Leu Val Thr Pro Ser Ala Phe Glu Ser Ala Glu Lys Thr Asp Val

195 200 205

Ile Pro Glu Trp Leu Lys Asn

210 215

<210> 69

<211> 648

<212> DNA

<213> Rabbit

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 69

atgaagcttc tcatcctcac ttgccttgtg gctactgctc ttgccaggca taaatttcat 60

ttaggacacc tgaaactcac tcaggagcag cctgagagca gtgagcagga aatcttaaaa 120

gaaagaaagc tcctcaggtt tgtccagaca gtaccactag aattaagaga ggaatatgtc 180

aatgaactga acaggcagag agaacttctg agagaaaaag agaatgagga aatcaaggga 240

actagaaatg aagtaactga ggaacatgtt ttggcagacc gtgagacaga agctagcatc 300

agctcatcaa gtgaggaaat tgttcccagc agcaccaagc agaagtacgt gccaagggaa 360

gacctggctt accaacctta cgtgcagcag cagcttctca gaatgaaaga acgctaccaa 420

atccaggaga gagagcctat gagagtggtg aatcaggaac tggctcagct ctatcttcag 480

cctttcgaac aaccctacca gcttgatgcc tatctccctg ctccttggta ctatactccg 540

gaagtgatgc agtatgttct ttccccactg ttctatgacc tcgttacacc cagtgccttt 600

gagagtgctg aaaaaactga cgttattcca gagtggttga agaattaa 648

<210> 70

<211> 214

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 70

Met Lys Leu Leu Ile Leu Thr Cys Leu Val Ala Val Ala Leu Ala Arg

1 5 10 15

Pro Lys His Pro Ile Lys His Gln Gly Leu Ser Pro Glu Val Leu Asn

20 25 30

Glu Asn Leu Leu Arg Phe Val Val Ala Pro Phe Pro Glu Val Phe Arg

35 40 45

Lys Glu Asn Ile Asn Glu Leu Ser Lys Asp Ile Gly Ser Glu Ser Ile

50 55 60

Glu Asp Gln Ala Met Glu Asp Ala Lys Gln Met Lys Ala Gly Ser Ser

65 70 75 80

Ser Ser Ser Glu Glu Ile Val Pro Asn Ser Ala Glu Gln Lys Tyr Ile

85 90 95

Gln Lys Glu Asp Val Pro Ser Glu Arg Tyr Leu Gly Tyr Leu Glu Gln

100 105 110

Leu Leu Arg Leu Lys Lys Tyr Asn Val Pro Gln Leu Glu Ile Val Pro

115 120 125

Lys Ser Ala Glu Glu Gln Leu His Ser Met Lys Glu Gly Asn Pro Ala

130 135 140

His Gln Lys Gln Pro Met Ile Ala Val Asn Gln Glu Leu Ala Tyr Phe

145 150 155 160

Tyr Pro Gln Leu Phe Arg Gln Phe Tyr Gln Leu Asp Ala Tyr Pro Ser

165 170 175

Gly Ala Trp Tyr Tyr Leu Pro Leu Gly Thr Gln Tyr Thr Asp Ala Pro

180 185 190

Ser Phe Ser Asp Ile Pro Asn Pro Ile Gly Ser Glu Asn Ser Gly Lys

195 200 205

Ile Thr Met Pro Leu Trp

210

<210> 71

<211> 645

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 71

atgaaacttc tcatccttac ctgtcttgtg gctgttgctc ttgccaggcc taaacatcct 60

atcaagcacc aaggactctc tccagaagtc ctcaatgaaa atttactcag gtttgttgtg 120

gcgccttttc cagaagtgtt tagaaaggag aacatcaatg aactgagtaa ggatattggg 180

agtgaatcaa ttgaggatca agccatggaa gatgctaagc aaatgaaagc tggaagcagt 240

tcgtcaagtg aggaaattgt tcccaatagt gctgagcaga agtacattca aaaggaagat 300

gtgccctctg agcgttacct gggttatctg gaacagcttc tcagactgaa aaaatacaac 360

gtgccccagc tggaaattgt tcccaaaagt gctgaggaac aacttcatag tatgaaagag 420

ggaaaccctg cccaccagaa acagcctatg atagcagtga atcaggaact ggcctacttc 480

taccctcagc ttttcagaca attctaccag ctggacgcct atccatctgg tgcctggtat 540

taccttccac taggcacaca atacactgat gccccctcat tctctgacat ccctaatccc 600

attggctctg agaacagtgg aaagattact atgccactgt ggtga 645

<210> 72

<211> 199

<212> PRT

<213> sperm whale

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 72

Val Leu Thr Thr Met Lys Leu Leu Ile Leu Thr Cys Leu Val Thr Ala

1 5 10 15

Ala Leu Ala Lys Pro Asn Glu Pro Asp Ser Arg Glu Pro Leu Ser Glu

20 25 30

Gln Leu Arg Glu Asp His Gly Met Glu Asp Pro Glu Gln Arg Gly Ser

35 40 45

Gly Ser Ser Ser Ser Ser Glu Val Asn Arg Phe Asp Val Asn Ser Ala

50 55 60

Ser Gln Leu Glu Asn Val Cys Glu Ser Leu Leu Tyr Gln Glu Gly Tyr

65 70 75 80

Leu Ser Pro Lys Glu Val Val Pro Asn Gly Asn Glu Arg Lys His Ile

85 90 95

Gln Gly Glu Asp Val Pro Ser Glu Arg Tyr Leu Arg Glu Pro Met Arg

100 105 110

Val Val Asn Gln Pro Phe Gln Gln Phe Tyr Gln Leu Asp Val His Pro

115 120 125

Tyr Ala Ala Trp Tyr Tyr Pro Pro Gln Val Thr Gln Tyr Ile Ala Ser

130 135 140

Pro Ser Phe Phe Asp Ile Pro Lys Pro Ile Ala Ser Glu Asn Gly Gly

145 150 155 160

Lys Thr Ile Met Pro Gln Trp Cys Phe Tyr His Met Ser Val Pro Asn

165 170 175

Glu Ser Thr Glu His Ser Phe Thr Cys Cys Glu Ser Tyr Arg Asn Lys

180 185 190

Ala Ile Asp Val Met Asn Ala

195

<210> 73

<211> 600

<212> DNA

<213> sperm whale

<220>

<221> misc_feature

<222> (0)...(0)

<223> partial cDNA of Casein alpha-S1

<400> 73

gtcttgacaa ccatgaaact tctcatcctt acctgccttg tgactgctgc tcttgccaag 60

cctaatgaac cagatagcag agagccttta tcagaacagc ttagagagga tcatggcatg 120

gaagaccctg agcaaagggg gtctggcagc agttcatcaa gtgaggtaaa tcgttttgat 180

gttaattcag catcccaatt agaaaatgtt tgtgaaagct tgttgtacca ggaaggttac 240

ctgtccccta aggaagttgt tcccaatggc aatgagcgga agcacattca aggggaagat 300

gtgccctctg aacgctatct gagagagcct atgagagtag tgaatcaacc tttccaacaa 360

ttctatcagc ttgatgtcca tccttatgct gcttggtatt atcctccaca agtcacacaa 420

tatattgctt ctccatcatt cttcgacatc cctaaaccca ttgcctctga gaacggtgga 480

aaaacaatta tgccacagtg gtgcttctat catatgtcgg ttcccaacga atcaacagaa 540

cacagcttca cgtgttgtga atcctacagg aataaagcca tcgatgtaat gaatgcatga 600

<210> 74

<211> 183

<212> PRT

<213> Australia needle mole

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 74

Met Lys Val Leu Ile Leu Ala Cys Leu Val Ala Phe Val Val Ala Met

1 5 10 15

Pro Glu Ser Pro Ser Ser Ser Ser Ser Ser Glu Glu Ala Ser Lys Ile

20 25 30

Leu Thr Lys Lys Arg Val Gln Arg Asp Gln Glu Tyr Tyr Leu Pro His

35 40 45

Gln Glu Glu Ser Val Ser Ser Ser Ser Ser Glu Glu Ser Thr Asp Arg

50 55 60

Leu Lys Arg Arg Leu Leu Lys Asp Lys Pro Ile Phe Arg Leu Leu Lys

65 70 75 80

Ala Thr Glu Ser Ser Ser Ser Glu Glu Ser Asp Ser Ala Ile Glu Lys

85 90 95

Arg Ile Leu Arg Glu Arg Gln Tyr Tyr Gln Gln Lys Leu Asp Glu Leu

100 105 110

Lys Glu Tyr Phe Arg Gln Phe Glu Pro Tyr Phe Tyr Pro Val Ala Tyr

115 120 125

Gln Lys Lys Glu Val Met Pro Tyr Gln Leu Glu Tyr Phe Val Pro Gln

130 135 140

Pro Glu Val Tyr Ser Ile Pro Gln Pro Val Tyr Arg Val Pro Gln Glu

145 150 155 160

Val Thr Phe Pro Ser Leu Leu His Phe Arg Tyr Ala Phe Pro Gln Ser

165 170 175

Thr Leu Pro Ile Glu Arg Lys

180

<210> 75

<211> 552

<212> DNA

<213> Australia needle mole

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 75

atgaaggtcc tcatcctggc ctgcctggtg gcttttgtcg tggcaatgcc tgagtcaccc 60

agcagcagtt catccagcga ggaagcttcc aaaattctga caaaaaagcg tgtccaaaga 120

gaccaagaat actaccttcc ccatcaggag gaatccgtaa gcagctcatc cagtgaggaa 180

tcgaccgatc gactcaaaag acgtctcctg aaagacaaac ccatcttccg tctcctgaag 240

gctacagaga gctcttcaag tgaggaatct gacagtgcta ttgaaaagcg tattctgagg 300

gagagacagt actatcaaca gaagctggat gaactcaaag aatattttcg tcagtttgaa 360

ccctacttct accctgtggc ttatcagaaa aaagaagtca tgccctacca gctggaatac 420

tttgttcctc aaccagaagt gtacagcatc ccccaaccag tatatagggt tcctcaagag 480

gtaaccttcc ccagtcttct ccatttccgc tacgcttttc ctcagtcaac cctccccatt 540

gagaggaagt aa 552

<210> 76

<211> 230

<212> PRT

<213> panda

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S1

<400> 76

Met Glu Ile Lys His Cys Pro Lys Asp Leu Lys Asp Gly Ser Leu Lys

1 5 10 15

Gln Cys Thr Gly Ser Gly His Lys Ala Ala Leu Ser Gln Glu Glu Gln

20 25 30

Gly Ser Ser Ala Val Ile Glu His Gly Ser Ile Pro Arg Gly Gly Asn

35 40 45

Asn Val His Arg Val Gly Gly Val Arg Ala Ser Cys Thr Leu Glu Ser

50 55 60

Thr Arg Arg Leu Leu Trp Met Cys Pro Ser Glu Lys Thr Ser Val Leu

65 70 75 80

Ala Leu Thr Thr Met Lys Val Leu Ile Leu Thr Cys Leu Val Ala Val

85 90 95

Ala Leu Ala Arg Pro Gly Gln Ala Val Glu Asp Pro Glu Gln Arg Gln

100 105 110

Ser Ser Ser Ser Ser Ser Ser Glu Glu Val Val Pro Ser Thr Thr Glu

115 120 125

Gln Lys Gln Ile Pro Arg Glu Asp Ile Leu Asn Gln Arg Tyr Leu Glu

130 135 140

Gln Leu Arg Arg Leu Ser Lys Tyr Asn Gln Gln Gln Gln Glu Thr Ile

145 150 155 160

His Asp Gln Gln Gln Leu Arg Gly Val Asn Glu Asn Asn Leu Leu Gln

165 170 175

Leu Pro Phe Gln Gln Phe Tyr Gln Leu Asp Ala Tyr Pro Phe Ala Ala

180 185 190

Trp Tyr Tyr Leu Pro Gln Ile Met Gln Tyr Ile Gly Tyr Thr Pro Ser

195 200 205

Tyr Asp Ile Ile Lys Pro Ile Ala Ser Glu Asn Ile Glu Asn Val Asp

210 215 220

Val Val Pro Glu Trp Trp

225 230

<210> 77

<211> 693

<212> DNA

<213> panda

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1 cDNA

<400> 77

atggaaatta aacactgtcc aaaagacctc aaagacgggt cacttaaaca gtgcacggga 60

agtggtcaca aggcagcgct aagccaggaa gagcagggaa gtagtgctgt aatagaacac 120

ggaagcattc cacgtggagg aaataacgtg catagggttg gaggagtaag agcttcatgc 180

actcttgaaa gcacaagaag attgttgtgg atgtgtcctt cagaaaagac gagtgtgctg 240

gccttgacaa ccatgaaggt tctcatcctt acctgccttg tggctgttgc tcttgccagg 300

cctggacagg ccgtggaaga ccctgagcaa aggcaatcta gcagcagttc gtcaagtgag 360

gaagttgttc ccagtaccac tgagcagaag caaattccaa gagaagacat actcaaccaa 420

cgctatctgg aacagcttcg tagactgagc aaatacaacc aacaacagca ggaaactatc 480

catgaccagc aacaacttcg cggagtgaat gaaaacaacc tcctccaact gcctttccaa 540

cagttctacc agcttgatgc ttatcccttt gctgcttggt attatcttcc acaaatcatg 600

cagtatattg gttatacacc atcctatgac atcattaaac ccattgcctc tgagaacatt 660

gaaaacgttg atgttgtgcc agagtggtgg taa 693

<210> 78

<211> 222

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 78

Met Lys Phe Phe Ile Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

1 5 10 15

Asn Thr Met Glu His Val Ser Ser Ser Glu Glu Ser Ile Ile Ser Gln

20 25 30

Glu Thr Tyr Lys Gln Glu Lys Asn Met Asp Ile Asn Pro Ser Lys Glu

35 40 45

Asn Leu Cys Ser Thr Phe Cys Lys Glu Val Val Arg Asn Ala Asn Glu

50 55 60

Glu Glu Tyr Ser Ile Gly Ser Ser Ser Glu Glu Ser Ala Glu Val Ala

65 70 75 80

Thr Glu Glu Val Lys Ile Thr Val Asp Asp Lys His Tyr Gln Lys Ala

85 90 95

Leu Asn Glu Ile Asn Gln Phe Tyr Arg Lys Phe Pro Gln Tyr Leu Gln

100 105 110

Tyr Leu Tyr Gln Gly Pro Ile Val Leu Asn Pro Trp Asp Gln Val Lys

115 120 125

Arg Asn Ala Val Pro Ile Thr Pro Thr Leu Asn Arg Glu Gln Leu Ser

130 135 140

Thr Ser Glu Glu Asn Ser Lys Lys Thr Val Asp Met Glu Ser Thr Glu

145 150 155 160

Val Phe Thr Lys Lys Thr Lys Leu Thr Glu Glu Glu Lys Asn Arg Leu

165 170 175

Asn Phe Leu Lys Lys Ile Ser Gln Arg Tyr Gln Lys Phe Ala Leu Pro

180 185 190

Gln Tyr Leu Lys Thr Val Tyr Gln His Gln Lys Ala Met Lys Pro Trp

195 200 205

Ile Gln Pro Lys Thr Lys Val Ile Pro Tyr Val Arg Tyr Leu

210 215 220

<210> 79

<211> 669

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 79

atgaagttct tcatctttac ctgccttttg gctgttgccc ttgcaaagaa tacgatggaa 60

catgtctcct ccagtgagga atctatcatc tcccaggaaa catataagca ggaaaagaat 120

atggacatta atcccagcaa ggagaacctt tgctccacat tctgcaagga agttgtaagg 180

aacgcaaatg aagaggaata ttctatcggc tcatctagtg aggaatctgc tgaagttgcc 240

acagaggaag ttaagattac tgtggacgat aagcactacc agaaagcact gaatgaaatc 300

aatcagtttt atcggaagtt cccccagtat ctccagtatc tgtatcaagg tccaattgtt 360

ttgaacccat gggatcaggt taagagaaat gctgttccca ttactcccac tctgaacaga 420

gagcagctct ccaccagtga ggaaaattca aagaagaccg ttgacatgga atcaacagaa 480

gtattcacta agaaaactaa actgactgaa gaagaaaaga atcgcctaaa ttttctgaaa 540

aaaatcagcc agcgttacca gaaattcgcc ttgccccagt atctcaaaac tgtttatcag 600

catcagaaag ctatgaagcc atggattcaa cctaagacaa aggttattcc ctatgtgagg 660

tacctttaa 669

<210> 80

<211> 223

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 80

Met Lys Phe Phe Ile Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

1 5 10 15

His Lys Met Glu His Val Ser Ser Ser Glu Glu Pro Ile Asn Ile Phe

20 25 30

Gln Glu Ile Tyr Lys Gln Glu Lys Asn Met Ala Ile His Pro Arg Lys

35 40 45

Glu Lys Leu Cys Thr Thr Ser Cys Glu Glu Val Val Arg Asn Ala Asn

50 55 60

Glu Glu Glu Tyr Ser Ile Arg Ser Ser Ser Glu Glu Ser Ala Glu Val

65 70 75 80

Ala Pro Glu Glu Ile Lys Ile Thr Val Asp Asp Lys His Tyr Gln Lys

85 90 95

Ala Leu Asn Glu Ile Asn Gln Phe Tyr Gln Lys Phe Pro Gln Tyr Leu

100 105 110

Gln Tyr Pro Tyr Gln Gly Pro Ile Val Leu Asn Pro Trp Asp Gln Val

115 120 125

Lys Arg Asn Ala Gly Pro Phe Thr Pro Thr Val Asn Arg Glu Gln Leu

130 135 140

Ser Thr Ser Glu Glu Asn Ser Lys Lys Thr Ile Asp Met Glu Ser Thr

145 150 155 160

Glu Val Phe Thr Lys Lys Thr Lys Leu Thr Glu Glu Glu Lys Asn Arg

165 170 175

Leu Asn Phe Leu Lys Lys Ile Ser Gln Tyr Tyr Gln Lys Phe Ala Trp

180 185 190

Pro Gln Tyr Leu Lys Thr Val Asp Gln His Gln Lys Ala Met Lys Pro

195 200 205

Trp Thr Gln Pro Lys Thr Asn Ala Ile Pro Tyr Val Arg Tyr Leu

210 215 220

<210> 81

<211> 672

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 81

atgaagttct tcatttttac ctgccttttg gccgttgccc ttgcaaagca taagatggaa 60

catgtctcct ccagtgagga acctatcaat atcttccagg aaatatataa gcaggaaaag 120

aatatggcca ttcatcccag aaaggagaaa ctttgcacca catcctgtga ggaagttgta 180

aggaacgcaa atgaagagga atattctatc agatcatcta gtgaggaatc tgctgaagtt 240

gccccagagg aaattaagat tactgtggac gataagcact accagaaagc cctgaatgaa 300

atcaatcagt tttatcagaa gttcccccag tatctccagt atccgtatca aggtccaatt 360

gttttgaacc catgggatca ggttaagaga aatgctggcc cctttactcc caccgtgaac 420

agagagcagc tctccaccag tgaggaaaat tcaaagaaga ccattgatat ggaatcaaca 480

gaagtattca ctaagaaaac taaactgact gaagaagaaa agaatcgcct aaattttctg 540

aaaaaaatca gccagtatta ccagaaattt gcctggcccc agtatctcaa gactgttgat 600

cagcatcaga aagctatgaa gccatggact caacctaaga caaatgctat tccctatgtg 660

aggtaccttt aa 672

<210> 82

<211> 239

<212> PRT

<213> domestic cat

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 82

Met Ser Gly Ala Glu Ala Val Asp Trp Glu Val Ser Glu Lys Lys Leu

1 5 10 15

His Ile Cys Arg Leu Ser Arg Gly Cys Gly Val Ser Pro Arg Val Thr

20 25 30

Met Asn Phe Leu Leu Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

35 40 45

His Glu Leu Lys Gln Leu Ser Ser Ser Glu Glu Ser Ala Thr Ser Ser

50 55 60

Ser Ser Gln Glu Thr Gly Asp Val Pro Thr Asp Thr Ile Glu Leu Thr

65 70 75 80

Glu Glu Glu Lys Val Tyr Leu Asn Gln Leu Ser Lys Ile Asn Gln Phe

85 90 95

Tyr Gln Ala Trp Asn Leu Pro Gln Tyr Leu Glu Ala Tyr His Gln Gln

100 105 110

His Ser Val Arg Asn Pro Trp Asn His Ile Lys Thr Asn Gly Tyr His

115 120 125

Leu Phe Pro Leu Leu Glu Lys Glu Tyr Leu Ser Ser Ser Glu Val Arg

130 135 140

Gly Phe Pro Val Arg Thr Glu Thr Trp His Pro Glu Thr Glu Ile Lys

145 150 155 160

Glu Val Gln Leu Asn Asp Glu Glu Lys Asn Tyr Leu Lys Gln Leu Val

165 170 175

Lys Ile Asn Gln Tyr Gln Gln Lys Phe Thr Phe Pro His Tyr Phe Gln

180 185 190

Ala Val His Pro Gln Gln Ile Ala Leu Asn Pro Trp Asn Arg Leu Lys

195 200 205

Glu Asn Thr Tyr Pro Phe Ile Leu Thr Leu Leu Gly Ser Leu Asn Gln

210 215 220

Phe Gly Pro Pro Asp Asp Glu Lys Gly Leu Asp Tyr Phe Ser Lys

225 230 235

<210> 83

<211> 720

<212> DNA

<213> domestic cat

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 83

atgtccggtg cagaggcagt ggactgggaa gttagtgaga agaagctgca catatgtcgt 60

ctttcaagag gctgcggggt cagtccccga gtaaccatga atttcctcct ctttacctgc 120

cttttggctg tggctctggc aaagcatgag ctaaagcagc tctcctccag cgaggaatct 180

gccaccagtt catctagtca ggaaactggt gatgtcccta ctgacacaat tgagctgact 240

gaggaagaaa aggtctacct caatcaactg agcaaaatca accagtttta tcaggcatgg 300

aatcttcccc agtatcttga ggcttatcat caacagcaca gtgttaggaa cccatggaat 360

cacattaaga caaatggcta tcatctattt ccccttctgg aaaaagagta cctgtcctcc 420

agtgaggtga ggggctttcc tgtgaggaca gaaacatggc atccagaaac agagattaag 480

gaagttcagc tgaatgatga agaaaagaat tacctgaaac agctggtcaa aatcaaccag 540

tatcaacaga agttcacctt cccccactat ttccaggctg ttcatccaca gcagatagct 600

ctgaaccctt ggaatcgcct taaggaaaat acttacccat tcattctcac tttgcttggt 660

tctctgaacc agtttggtcc tcctgatgat gaaaagggat tagactactt ttctaaataa 720

<210> 84

<211> 121

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 84

Met Lys Phe Phe Ile Phe Thr Cys Leu Leu Ala Ile Ala Leu Ala Lys

1 5 10 15

His Glu Ser Ala Glu Val Ser Thr Glu Glu Val Lys His Thr Val Asp

20 25 30

Gln Lys His Tyr Val Lys Gln Leu Asn Lys Ile Asn Pro Phe Tyr Gln

35 40 45

Lys Trp Asn Phe Leu Pro Phe Leu Gln Lys Thr Glu Leu Thr Glu Glu

50 55 60

Glu Lys Asn Asp Gln Lys His Leu Asn Lys Ile Asn Gln Tyr Tyr Gln

65 70 75 80

Phe Thr Leu Pro Gln Tyr Val Lys Ala Val Tyr Gln Tyr His Lys Ile

85 90 95

Met Lys Pro Trp Lys Asn Met Lys Thr Asn Ala Tyr Gln Val Ile Pro

100 105 110

Thr Leu Gly Ser Leu Arg Phe Leu Asn

115 120

<210> 85

<211> 84

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2

<400> 85

atgaagttct tcatctttac ctgccttttg gctgttgctc tggcacatca tgagataaag 60

cactcctcct cttccagtga gtaa 84

<210> 86

<211> 182

<212> PRT

<213> Rabbit

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 86

Met Lys Phe Phe Ile Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

1 5 10 15

Pro Lys Ile Glu Gln Ser Ser Ser Glu Glu Thr Ile Ala Val Ser Gln

20 25 30

Glu Val Ser Pro Asn Leu Glu Asn Ile Cys Ser Thr Ala Cys Glu Glu

35 40 45

Pro Ile Lys Asn Ile Asn Glu Val Glu Tyr Val Glu Val Pro Thr Glu

50 55 60

Ile Lys Asp Gln Glu Phe Tyr Gln Lys Val Asn Leu Leu Gln Tyr Leu

65 70 75 80

Gln Ala Leu Tyr Gln Tyr Pro Thr Val Met Asp Pro Trp Thr Arg Ala

85 90 95

Glu Thr Lys Ala Ile Pro Phe Ile Arg Thr Met Gln Tyr Lys Gln Glu

100 105 110

Lys Asp Ala Thr Lys His Thr Ser Gln Lys Thr Glu Leu Thr Glu Glu

115 120 125

Glu Lys Ala Phe Leu Lys Tyr Leu Asp Glu Met Lys Gln Tyr Tyr Gln

130 135 140

Lys Phe Val Phe Pro Gln Tyr Leu Lys Asn Ala His His Phe Gln Lys

145 150 155 160

Thr Met Asn Pro Trp Asn His Val Lys Thr Ile Ile Tyr Gln Ser Val

165 170 175

Pro Thr Leu Arg Tyr Leu

180

<210> 87

<211> 549

<212> DNA

<213> Rabbit

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 87

atgaagttct tcattttcac ctgccttctg gctgttgctc tggcaaagcc taagattgag 60

caatcttcaa gtgaggaaac tattgctgtc tcccaagaag tatccccaaa cttagaaaac 120

atttgttcta cagcctgtga ggaacccata aagaatatca atgaagtgga atacgttgaa 180

gttcccacag agataaaaga tcaggaattt tatcagaagg tgaacctcct ccagtatctc 240

caggctcttt accaatatcc cactgtcatg gacccatgga ctcgggctga gacaaaggcc 300

atccccttta ttcgcactat gcaatataag caggaaaagg atgccactaa gcataccagt 360

cagaaaaccg aactgaccga agaagaaaag gcttttctaa aatacttgga tgaaatgaaa 420

caatattatc agaagttcgt ttttccccaa tacctaaaaa atgctcatca ttttcagaaa 480

actatgaacc cttggaatca tgttaagaca attatttacc aaagtgtgcc cactctgaga 540

tacttataa 549

<210> 88

<211> 223

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 88

Met Lys Phe Phe Ile Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

1 5 10 15

His Lys Met Glu His Val Ser Ser Ser Glu Glu Pro Ile Asn Ile Ser

20 25 30

Gln Glu Ile Tyr Lys Gln Glu Lys Asn Met Ala Ile His Pro Arg Lys

35 40 45

Glu Lys Leu Cys Thr Thr Ser Cys Glu Glu Val Val Arg Asn Ala Asp

50 55 60

Glu Glu Glu Tyr Ser Ile Arg Ser Ser Ser Glu Glu Ser Ala Glu Val

65 70 75 80

Ala Pro Glu Glu Val Lys Ile Thr Val Asp Asp Lys His Tyr Gln Lys

85 90 95

Ala Leu Asn Glu Ile Asn Gln Phe Tyr Gln Lys Phe Pro Gln Tyr Leu

100 105 110

Gln Tyr Leu Tyr Gln Gly Pro Ile Val Leu Asn Pro Trp Asp Gln Val

115 120 125

Lys Arg Asn Ala Gly Pro Phe Thr Pro Thr Val Asn Arg Glu Gln Leu

130 135 140

Ser Thr Ser Glu Glu Asn Ser Lys Lys Thr Ile Asp Met Glu Ser Thr

145 150 155 160

Glu Val Phe Thr Lys Lys Thr Lys Leu Thr Glu Glu Glu Lys Asn Arg

165 170 175

Leu Asn Phe Leu Lys Lys Ile Ser Gln Tyr Tyr Gln Lys Phe Ala Trp

180 185 190

Pro Gln Tyr Leu Lys Thr Val Asp Gln His Gln Lys Ala Met Lys Pro

195 200 205

Trp Thr Gln Pro Lys Thr Asn Ala Ile Pro Tyr Val Arg Tyr Leu

210 215 220

<210> 89

<211> 672

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 89

atgaagttct tcatttttac ctgccttttg gccgttgccc ttgcaaagca taagatggaa 60

catgtctcct ccagtgagga acctatcaat atctcccagg aaatatataa gcaggaaaag 120

aatatggcca ttcatcccag aaaggagaaa ctttgcacca catcctgtga ggaagttgta 180

aggaacgcag atgaagagga atattctatc agatcatcta gtgaggaatc tgctgaagtt 240

gccccagagg aagttaagat tactgtggac gataagcact accagaaagc actgaatgaa 300

atcaatcagt tttatcagaa gttcccccag tacctccagt atctgtatca aggcccaatt 360

gttttgaacc catgggatca ggttaagaga aatgctggcc cctttactcc caccgtgaac 420

agagagcagc tctccaccag tgaggaaaat tcaaagaaga ccattgatat ggaatcaaca 480

gaagtattca ctaagaaaac taaactgact gaagaagaaa agaatcgcct aaattttctg 540

aaaaaaatca gccagtatta ccagaaattt gcctggcccc agtatctcaa gactgttgat 600

cagcatcaga aagctatgaa gccatggact caacctaaga caaacgctat tccctatgtg 660

aggtaccttt aa 672

<210> 90

<211> 243

<212> PRT

<213> sperm whale

<220>

<221> variants

<222> (0)...(0)

<223> Casein alpha-S2

<400> 90

Met Lys Phe Phe Leu Phe Thr Cys Leu Leu Ala Val Ala Leu Ala Lys

1 5 10 15

His Glu Met Glu His Val Ser Ser Ser Glu Glu Ser Ile Asn Met Phe

20 25 30

Gln Glu Lys Tyr Lys Gln Arg Lys Asn Val Val Leu His Pro Ser Lys

35 40 45

Glu Asn Ile Cys Ser Thr Ser Cys Glu Val Cys Ile Asp Phe Thr Ser

50 55 60

Val Val Gln Cys Glu Val Lys Tyr Ser Ile Arg Ser Ser Pro Lys Glu

65 70 75 80

Thr Ala Glu Val Pro Arg Glu Lys Val Lys Leu Thr Val Glu Asp Lys

85 90 95

Gln Tyr Leu Lys Gln Leu Ser Lys Ile Ser Gln Phe Tyr Gln Lys Phe

100 105 110

Pro Gln Tyr Ile Gln Ala Leu Tyr Gln Ala Pro Thr Val Met Asn Pro

115 120 125

Trp Gly Gln Val Lys Arg Ser Ala Glu Pro Phe Ile Leu Thr Val Ser

130 135 140

Arg Gln Gln Leu Ser Thr Gly Glu Glu Asn Ser Lys Lys Thr Val Asp

145 150 155 160

Met Glu Ser Thr Glu Val Leu Thr Lys Lys Thr Thr Leu Thr Glu Glu

165 170 175

Glu Lys Asn Arg Leu Lys Phe Leu Asn Lys Ile Asn Gln Tyr Tyr Gln

180 185 190

Lys Leu Thr Trp Pro Gln Tyr Leu Lys Thr Ile Ser Gln Tyr Gln Lys

195 200 205

Thr Val Lys Pro Trp Asn His Ile Lys Thr Asn Val Ile Pro Tyr Leu

210 215 220

Asp Tyr Ser Gly Asn Pro Gly Ser Pro Tyr Pro Val Asn Pro Thr Leu

225 230 235 240

Asn Ile Pro

<210> 91

<211> 732

<212> DNA

<213> sperm whale

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2 cDNA

<400> 91

atgaagttct tcctttttac ctgccttttg gctgttgctc ttgcaaagca tgagatggaa 60

catgtctcct ccagtgagga atctatcaac atgttccagg aaaaatataa gcagagaaag 120

aatgtggtcc ttcatcccag caaggagaac atttgttcca catcctgtga ggtatgtatt 180

gatttcacat cagtagtaca atgtgaagta aaatatagta ttagatcttc ccctaaggaa 240

actgctgaag ttcccaggga gaaagttaag cttactgtgg aagataagca atacctgaaa 300

caactgagca aaatcagtca gttttatcag aagttccccc aatatatcca ggctctatat 360

caagctccaa ctgttatgaa cccatggggt caagttaaga gaagtgctga gccctttatt 420

ctcactgtga gcagacagca gctctctacc ggtgaggaaa attcaaagaa gactgttgat 480

atggaatcaa cggaagtact cactaagaaa actacattga ctgaagaaga aaagaatcgc 540

ctaaaatttc tgaacaaaat caaccaatat tatcagaaat taacctggcc ccagtatctc 600

aagactattt ctcagtatca gaaaactgtg aagccgtgga atcacattaa gacaaatgtt 660

atcccctatc tggattacag cggcaatcct ggcagtcctt acccagttaa cccaactctg 720

aatattcctt aa 732

<210> 92

<211> 142

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 92

Met Met Ser Phe Val Ser Leu Leu Leu Val Gly Ile Leu Phe His Ala

1 5 10 15

Thr Gln Ala Glu Gln Leu Thr Lys Cys Glu Val Phe Arg Glu Leu Lys

20 25 30

Asp Leu Lys Gly Tyr Gly Gly Val Ser Leu Pro Glu Trp Val Cys Thr

35 40 45

Thr Phe His Thr Ser Gly Tyr Asp Thr Gln Ala Ile Val Gln Asn Asn

50 55 60

Asp Ser Thr Glu Tyr Gly Leu Phe Gln Ile Asn Asn Lys Ile Trp Cys

65 70 75 80

Lys Asp Asp Gln Asn Pro His Ser Ser Asn Ile Cys Asn Ile Ser Cys

85 90 95

Asp Lys Phe Leu Asp Asp Asp Leu Thr Asp Asp Ile Met Cys Val Lys

100 105 110

Lys Ile Leu Asp Lys Val Gly Ile Asn Tyr Trp Leu Ala His Lys Ala

115 120 125

Leu Cys Ser Glu Lys Leu Asp Gln Trp Leu Cys Glu Lys Leu

130 135 140

<210> 93

<211> 429

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> alpha-lactalbumin cDNA

<400> 93

atgatgtcct ttgtctctct gctcctggta ggcatcctat tccatgccac ccaggctgaa 60

cagttaacaa aatgtgaggt gttccgggag ctgaaagact tgaagggcta cggaggtgtc 120

agtttgcctg aatgggtctg taccacgttt cataccagtg gttatgacac acaagccata 180

gtacaaaaca atgacagcac agaatatgga ctcttccaga taaataataa aatttggtgc 240

aaagacgacc agaaccctca ctcaagcaac atctgtaaca tctcctgtga caagttcctg 300

gatgatgatc ttactgatga cattatgtgt gtcaagaaga ttctggataa agtaggaatt 360

aactactggt tggcccataa agcactctgt tctgagaagc tggatcagtg gctctgtgag 420

aagttgtga 429

<210> 94

<211> 142

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 94

Met Met Ser Phe Val Ser Leu Leu Leu Val Gly Ile Leu Phe His Ala

1 5 10 15

Thr Gln Ala Glu Gln Leu Thr Lys Cys Glu Val Phe Gln Lys Leu Lys

20 25 30

Asp Leu Lys Asp Tyr Gly Gly Val Ser Leu Pro Glu Trp Val Cys Thr

35 40 45

Ala Phe His Thr Ser Gly Tyr Asp Thr Gln Ala Ile Val Gln Asn Asn

50 55 60

Asp Ser Thr Glu Tyr Gly Leu Phe Gln Ile Asn Asn Lys Ile Trp Cys

65 70 75 80

Lys Asp Asp Gln Asn Pro His Ser Arg Asn Ile Cys Asn Ile Ser Cys

85 90 95

Asp Lys Phe Leu Asp Asp Asp Leu Thr Asp Asp Ile Val Cys Ala Lys

100 105 110

Lys Ile Leu Asp Lys Val Gly Ile Asn Tyr Trp Leu Ala His Lys Ala

115 120 125

Leu Cys Ser Glu Lys Leu Asp Gln Trp Leu Cys Glu Lys Leu

130 135 140

<210> 95

<211> 735

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> alpha-lactalbumin cDNA

<400> 95

ggggggtaac caaaatgatg tcctttgtct ctctgctcct ggtaggcatc ctgttccacg 60

ccacccaggc tgaacaatta acaaaatgtg aggtgttcca gaagctgaag gacttgaagg 120

actacggagg tgtcagtttg cctgaatggg tctgtactgc atttcatacc agtggttatg 180

acacacaagc catagtacaa aacaatgaca gcacagaata tggactcttc cagataaata 240

ataaaatttg gtgcaaagac gaccagaacc ctcactcaag gaacatctgt aacatctcct 300

gtgacaagtt cctggatgat gatcttactg atgacattgt gtgtgccaag aagattctgg 360

ataaagtagg aattaactac tggttggccc ataaagcact ctgttctgag aagctggatc 420

agtggctctg tgagaagttg tgaacacctg ctgtctttgc tgcttctgtc ctctttctgt 480

tcctggaact cctctgcccc ttggctacct cgttttgctt ctttgtaccc ccttgaagct 540

aactcgtctc tgagccctgg gccctgtagt gacgatggac atgtaaggac taatctccag 600

ggatgcgtga atggtgctca ggacatttga cccttgctcg gtgcccctga tagcactttt 660

aatgcaacag tgcatattcc acttctgtcc tgaataaaag gcctgattct gaaaaaaaaa 720

aaaaaaaaaa aaaaa 735

<210> 96

<211> 142

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 96

Met Arg Phe Phe Val Pro Leu Phe Leu Val Gly Ile Leu Phe Pro Ala

1 5 10 15

Ile Leu Ala Lys Gln Phe Thr Lys Cys Glu Leu Ser Gln Leu Leu Lys

20 25 30

Asp Ile Asp Gly Tyr Gly Gly Ile Ala Leu Pro Glu Leu Ile Cys Thr

35 40 45

Met Phe His Thr Ser Gly Tyr Asp Thr Gln Ala Ile Val Glu Asn Asn

50 55 60

Glu Ser Thr Glu Tyr Gly Leu Phe Gln Ile Ser Asn Lys Leu Trp Cys

65 70 75 80

Lys Ser Ser Gln Val Pro Gln Ser Arg Asn Ile Cys Asp Ile Ser Cys

85 90 95

Asp Lys Phe Leu Asp Asp Asp Ile Thr Asp Asp Ile Met Cys Ala Lys

100 105 110

Lys Ile Leu Asp Ile Lys Gly Ile Asp Tyr Trp Leu Ala His Lys Ala

115 120 125

Leu Cys Thr Glu Lys Leu Glu Gln Trp Leu Cys Glu Lys Leu

130 135 140

<210> 97

<211> 701

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 97

atgaggttct ttgtccctct gttcctggtg ggcatcctgt tccctgccat cctggccaag 60

caattcacaa aatgtgagct gtcccagctg ctgaaagaca tagatggtta tggaggcatc 120

gctttgcctg aattgatctg taccatgttt cacaccagtg gttatgacac acaagccata 180

gttgaaaaca atgaaagcac ggaatatgga ctcttccaga tcagtaataa gctttggtgc 240

aagagcagcc aggtccctca gtcaaggaac atctgtgaca tctcctgtga caagttcctg 300

gatgatgaca ttactgatga cataatgtgt gccaagaaga tcctggatat taaaggaatt 360

gactactggt tggcccataa agccctctgc actgagaagc tggaacagtg gctttgtgag 420

aagttgtgag tgtctgctgt ccttggcacc cctgcccact ccacactcct ggaatacctc 480

ttccctaatg ccacctcagt ttgtttcttt ctgttccccc aaagcttatc tgtctctgag 540

ccttgggccc tgtagtgaca tcaccgaatt cttgaagact attttccagg gatgcctgag 600

tggtgcactg agctctagac ccttactcag tgccttcgat ggcactttca ctacagcaca 660

gatttcacct ctgtcttgaa taaaggtccc actttgaagt c 701

<210> 98

<211> 142

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 98

Met Met Ser Phe Val Ser Leu Leu Leu Val Gly Ile Leu Phe His Ala

1 5 10 15

Thr Gln Ala Glu Gln Leu Thr Lys Cys Glu Ala Phe Gln Lys Leu Lys

20 25 30

Asp Leu Lys Asp Tyr Gly Gly Val Ser Leu Pro Glu Trp Val Cys Thr

35 40 45

Ala Phe His Thr Ser Gly Tyr Asp Thr Gln Ala Ile Val Gln Asn Asn

50 55 60

Asp Ser Thr Glu Tyr Gly Leu Phe Gln Ile Asn Asn Lys Ile Trp Cys

65 70 75 80

Lys Asp Asp Gln Asn Pro His Ser Arg Asn Ile Cys Asn Ile Ser Cys

85 90 95

Asp Lys Phe Leu Asp Asp Asp Leu Thr Asp Asp Ile Val Cys Ala Lys

100 105 110

Lys Ile Leu Asp Lys Val Gly Ile Asn Tyr Trp Leu Ala His Lys Ala

115 120 125

Leu Cys Ser Glu Lys Leu Asp Gln Trp Leu Cys Glu Lys Leu

130 135 140

<210> 99

<211> 723

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 99

ttccaggatc ttagggggta accaaaatga tgtcctttgt ctctctgctc ctggtaggca 60

tcctattcca tgccacccag gctgaacaat taacaaaatg tgaggcgttc cagaagctga 120

aggacttgaa ggactacgga ggtgtcagtt tgcctgaatg ggtctgtacc gcatttcata 180

ccagtggtta tgacacacaa gccatagtac aaaacaatga cagcacagaa tatggactct 240

tccagataaa taataaaatt tggtgcaaag acgaccagaa ccctcactca aggaacatct 300

gtaacatctc ctgtgacaag ttcctggatg atgatcttac tgatgacatt gtgtgtgcca 360

agaagattct ggataaagta ggaattaact actggttggc ccataaagca ctctgttctg 420

agaagctgga tcagtggctc tgtgagaagt tgtgaacacc tgctgtcttt gctgcttctg 480

ccctctttct gttcctggaa ctcctctgcc ccttggctac ctcgttttgc ttctttgtac 540

ccccttgaag ctaacttgtc tctgagccct gggccctgta gtggcgatgg acatgtaagg 600

actaatctct agggatgcat gaatggtgct cgggacattt gacccttgct cggtgcccct 660

gatagcactt ttaatgcaac agtgcatatt ccacttctgt cctgaataaa aggcctgatt 720

ctg 723

<210> 100

<211> 178

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 100

Met Lys Cys Leu Leu Leu Ala Leu Ala Leu Thr Cys Gly Ala Gln Ala

1 5 10 15

Leu Ile Val Thr Gln Thr Met Lys Gly Leu Asp Ile Gln Lys Val Ala

20 25 30

Gly Thr Trp Tyr Ser Leu Ala Met Ala Ala Ser Asp Ile Ser Leu Leu

35 40 45

Asp Ala Gln Ser Ala Pro Leu Arg Val Tyr Val Glu Glu Leu Lys Pro

50 55 60

Thr Pro Glu Gly Asp Leu Glu Ile Leu Leu Gln Lys Trp Glu Asn Asp

65 70 75 80

Glu Cys Ala Gln Lys Lys Ile Ile Ala Glu Lys Thr Lys Ile Pro Ala

85 90 95

Val Phe Lys Ile Asp Ala Leu Asn Glu Asn Lys Val Leu Val Leu Asp

100 105 110

Thr Asp Tyr Lys Lys Tyr Leu Leu Val Cys Met Glu Asn Ser Ala Glu

115 120 125

Pro Glu Gln Ser Leu Val Cys Gln Cys Leu Val Arg Thr Pro Glu Val

130 135 140

Asp Asp Glu Ala Leu Glu Lys Phe Asp Lys Ala Leu Lys Ala Leu Pro

145 150 155 160

Met His Ile Arg Leu Ser Phe Asn Pro Thr Gln Leu Glu Glu Gln Cys

165 170 175

His Ile

<210> 101

<211> 790

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 101

actccactcc ctgcagagct cagaagcgtg atcccggctg cagccatgaa gtgcctcctg 60

cttgccctgg ccctcacctg tggcgcccag gccctcatcg tcacccagac catgaagggc 120

ctggatatcc agaaggtggc ggggacttgg tactccttgg ccatggcggc cagcgacatc 180

tccctgctgg acgcccagag tgcccccctg agagtgtatg tggaggagct gaagcccacc 240

cctgagggcg acctggagat cctgctgcag aaatgggaga atgatgagtg tgctcagaag 300

aagatcattg cagaaaaaac caagatccct gcggtgttca agatcgatgc cttgaacgag 360

aacaaagtcc ttgtgctgga caccgactac aaaaagtacc tgctcgtctg catggagaac 420

agtgctgagc ccgagcaaag cctggtctgc cagtgcctgg tcaggacccc ggaggtggac 480

gacgaggccc tggagaaatt cgacaaagcc ctcaaggccc tgcccatgca catccggctg 540

tccttcaacc caacccagct ggaggagcag tgccacatct aggtgagccc ctgccggcgc 600

ctctgggagc ccgggagcct tggcccctct ggggacagac gatgtcatcc ccgcctgccc 660

catcagggga ccaggaggaa ccgggaccac attcacccct cctgggaccc aggcccctcc 720

aggcccctcc tggggcctcc tgcttggggc cgctcctcct tcagcaataa aggcataaac 780

ctgtgctctc 790

<210> 102

<211> 180

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 102

Met Lys Cys Leu Leu Leu Ala Leu Gly Leu Ala Leu Ala Cys Gly Ile

1 5 10 15

Gln Ala Ile Ile Val Thr Gln Thr Met Lys Gly Leu Asp Ile Gln Lys

20 25 30

Val Ala Gly Thr Trp Tyr Ser Leu Ala Met Ala Ala Ser Asp Ile Ser

35 40 45

Leu Leu Asp Ala Gln Ser Ala Pro Leu Arg Val Tyr Val Glu Glu Leu

50 55 60

Lys Pro Thr Pro Glu Gly Asn Leu Glu Ile Leu Leu Gln Lys Trp Glu

65 70 75 80

Asn Gly Glu Cys Ala Gln Lys Lys Ile Ile Ala Glu Lys Thr Lys Ile

85 90 95

Pro Ala Val Phe Lys Ile Asp Ala Leu Asn Glu Asn Lys Val Leu Val

100 105 110

Leu Asp Thr Asp Tyr Lys Lys Tyr Leu Leu Phe Cys Met Glu Asn Ser

115 120 125

Ala Glu Pro Glu Gln Ser Leu Ala Cys Gln Cys Leu Val Arg Thr Pro

130 135 140

Glu Val Asp Lys Glu Ala Leu Glu Lys Phe Asp Lys Ala Leu Lys Ala

145 150 155 160

Leu Pro Met His Ile Arg Leu Ala Phe Asn Pro Thr Gln Leu Glu Gly

165 170 175

Gln Cys His Val

180

<210> 103

<211> 786

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 103

actccctgca gagctcagaa gcacgacccc agctgcagcc atgaagtgcc tcctgcttgc 60

cctgggcctg gccctcgcct gtggcatcca ggccatcatc gtcacccaga ccatgaaagg 120

cctggacatc cagaaggtgg cggggacttg gtactccttg gctatggcgg ccagcgacat 180

ctccctgctg gacgcccaga gtgcccccct gagagtgtac gtggaggagc tgaagcccac 240

ccccgagggc aacctggaga tcctgctgca gaaatgggag aacggtgagt gtgctcagaa 300

gaagattatt gcagaaaaaa ccaagatccc tgcggtgttc aagatcgatg ccttgaacga 360

gaacaaagtc cttgtgctgg acaccgacta caaaaagtac ctgctcttct gcatggaaaa 420

cagtgctgag cccgagcaaa gcctggcctg ccagtgcctg gtcaggaccc cagaggtgga 480

caaggaggcc ctggagaaat tcgacaaagc cctcaaggcc ctgcccatgc acatccggct 540

cgccttcaac ccgacccagc tggaggggca gtgccacgtc taggtgagcc cctgccggcg 600

cctctgtggg cccgggagcc ttggcccctc tggggacaga cgacgtcacc cccgcctccc 660

ccatcagggg gaccaggagg gaccgggacc acggtcacct ctcctgggac ccaggcccct 720

ccaggcccct cctgtggcct cctgctcggg gccgctcctc cttcagcaat aaaggcataa 780

acctgt 786

<210> 104

<211> 181

<212> PRT

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 104

Met Ala Leu Glu Lys Gly Pro Leu Leu Leu Leu Ala Leu Gly Leu Gly

1 5 10 15

Leu Ala Gly Ala Gln Lys Ala Leu Glu Glu Val Pro Val Gln Pro Gly

20 25 30

Phe Asn Ala Gln Lys Val Glu Gly Arg Trp Leu Thr Leu Gln Leu Ala

35 40 45

Ala Asn His Ala Asp Leu Val Ser Pro Ala Asp Pro Leu Arg Leu Ala

50 55 60

Leu His Ser Ile Arg Thr Arg Asp Gly Gly Asp Val Asp Phe Val Leu

65 70 75 80

Phe Trp Lys Gly Glu Gly Val Cys Lys Glu Thr Asn Ile Thr Val His

85 90 95

Pro Thr Gln Leu Gln Gly Gln Tyr Gln Gly Ser Phe Glu Gly Gly Ser

100 105 110

Met His Val Cys Phe Val Ser Thr Asp Tyr Ser Asn Leu Ile Leu Tyr

115 120 125

Val Arg Phe Glu Asp Asp Glu Ile Thr Asn Leu Trp Val Leu Leu Ala

130 135 140

Arg Arg Met Leu Glu Asp Pro Lys Trp Leu Gly Arg Tyr Leu Glu Tyr

145 150 155 160

Val Glu Lys Phe His Leu Gln Lys Ala Pro Val Phe Asn Ile Asp Gly

165 170 175

Pro Cys Pro Pro Pro

180

<210> 105

<211> 1505

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 105

tcccccttcc ccggagccat ggccctggag aaaggcccgc tcctgctgct ggcccttggc 60

ctgggcctgg cgggtgccca gaaggctctg gaagaggtac cggtacagcc gggcttcaat 120

gcgcagaagg tggaggggcg ctggctcacc ctgcagctgg cagccaacca cgcagacctg 180

gtctccccgg ccgaccccct gaggctcgct ctccactcca tccggaccag ggacggcggg 240

gacgtggact tcgtgctgtt ctggaaggga gaaggggtgt gtaaagaaac aaacatcacc 300

gtccatccaa cccagttgca aggccagtac caaggctcat tcgagggcgg cagcatgcac 360

gtatgcttcg tcagcaccga ctacagcaac ctcattcttt acgtgcgctt tgaggatgat 420

gagatcacca acctgtgggt gctgctggcg agaagaatgc tggaggaccc caaatggctg 480

ggaagatact tggagtacgt ggagaaattc cacctgcaga aagccccggt cttcaacata 540

gatggcccat gtcccccacc ctgagcctag gtctggcggt tctggagtct tcctgcctgg 600

gcccctcacc cctctgctgc cctcagcctc ccttccacct ccttcacctt ggcttgtggc 660

ctggactgtc cccaggtccc cctggaagcc cttttgcatc tcagggactc aaggaagctc 720

cccagctgag cccaaccctg cctctctcct ggtcccctcc cctgctggga aggcctcttc 780

cctctgtgcg tctccaggtc ctgccaacca cctgccaacc aacagccaag ggccagcagt 840

gtgccccagc ctggcctgtg ggcctggagc acacccaggg tggtgaggag ggggcacatg 900

gccccctgag ctcctgcccg cagcgccaga ggcctccaaa acttatacaa tgagtggagc 960

actgtagtcc caggtgcctc cgatgcaccc ccctccccag ggctgctggg gtggccctca 1020

agtgtccttc aggaacatga ccccacggag gctgttctca gactccagct cccctccact 1080

gtgacccacc tcacctgggt ctgctgggga ccctccagag aggtggcctc catgctccgt 1140

gagcaaacgc atatgtcccc actgaggtcc aagagcccta agtgagccca gctccagacc 1200

ctgctctctg cagaagccca gcggggctgc caggtaaaca cagacagctg tacctgtgtg 1260

gcaggtgaga ccagccaacg cccactcctc ggagcccagg attctgaagg gcggcgccca 1320

cttctgcacc cggtgagcca gggctgccca tcggcagggc aggctctgag gaaattgggt 1380

cagggactca atacttgtcg tctggaactc ccaagacagg tatgtccaga ggctgccccg 1440

aaaacgcctc cagtgaggcc ttctccccct tctccccctt ctcccccttc tccccaatct 1500

ccccc 1505

<210> 106

<211> 180

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 106

Met Lys Cys Leu Leu Leu Ala Leu Gly Leu Ala Leu Ala Cys Gly Val

1 5 10 15

Gln Ala Ile Ile Val Thr Gln Thr Met Lys Gly Leu Asp Ile Gln Lys

20 25 30

Val Ala Gly Thr Trp His Ser Leu Ala Met Ala Ala Ser Asp Ile Ser

35 40 45

Leu Leu Asp Ala Gln Ser Ala Pro Leu Arg Val Tyr Val Glu Glu Leu

50 55 60

Lys Pro Thr Pro Glu Gly Asn Leu Glu Ile Leu Leu Gln Lys Trp Glu

65 70 75 80

Asn Gly Glu Cys Ala Gln Lys Lys Ile Ile Ala Glu Lys Thr Lys Ile

85 90 95

Pro Ala Val Phe Lys Ile Asp Ala Leu Asn Glu Asn Lys Val Leu Val

100 105 110

Leu Asp Thr Asp Tyr Lys Lys Tyr Leu Leu Phe Cys Met Glu Asn Ser

115 120 125

Ala Glu Pro Glu Gln Ser Leu Ala Cys Gln Cys Leu Val Arg Thr Pro

130 135 140

Glu Val Asp Asn Glu Ala Leu Glu Lys Phe Asp Lys Ala Leu Lys Ala

145 150 155 160

Leu Pro Met His Ile Arg Leu Ala Phe Asn Pro Thr Gln Leu Glu Gly

165 170 175

Gln Cys His Val

180

<210> 107

<211> 785

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 107

ctccctgcag agttcagaag cacgacccca gctgcagcca tgaagtgcct cctgcttgcc 60

ctgggcctgg ccctcgcctg tggcgtccag gccatcatcg tcacccagac catgaaaggc 120

ctggacatcc agaaggtggc ggggacttgg cactccttgg ctatggcggc cagcgacatc 180

tccctgctgg atgcccagag tgcccccctg agagtgtacg tggaggagct gaagcccacc 240

cccgagggca acctggagat cctgctgcag aaatgggaga acggcgagtg tgctcagaag 300

aagattattg cagaaaaaac caagatccct gcggtgttca agatcgatgc cttgaatgag 360

aacaaagtcc ttgtgctgga caccgactac aaaaagtacc tgctcttctg catggaaaac 420

agtgctgagc ccgagcaaag cctggcctgc cagtgcctgg tcaggacccc ggaggtggac 480

aacgaggccc tggagaaatt cgacaaagcc ctcaaggccc tgcccatgca catccggctt 540

gccttcaacc cgacccagct ggaggggcag tgccacgtct aggtgagccc ctgccggtgc 600

ctctgggggc ccgggagcct tggctcctct ggggacagac gacgtcacca ccgccccccc 660

ccatcagggg gactagaagg gaccaggact gcagtcaccc ttcctgggac ccaggcccct 720

ccaggcccct cctggggctc ctgctctggg cagcttctcc ttcaccaata aaggcataaa 780

cctgt 785

<210> 108

<211> 708

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> lactoferrin

<400> 108

Met Lys Leu Phe Val Pro Ala Leu Leu Ser Leu Gly Ala Leu Gly Leu

1 5 10 15

Cys Leu Ala Ala Pro Arg Lys Asn Val Arg Trp Cys Thr Ile Ser Gln

20 25 30

Pro Glu Trp Phe Lys Cys Arg Arg Trp Gln Trp Arg Met Lys Lys Leu

35 40 45

Gly Ala Pro Ser Ile Thr Cys Val Arg Arg Ala Phe Ala Leu Glu Cys

50 55 60

Ile Arg Ala Ile Ala Glu Lys Lys Ala Asp Ala Val Thr Leu Asp Gly

65 70 75 80

Gly Met Val Phe Glu Ala Gly Arg Asp Pro Tyr Lys Leu Arg Pro Val

85 90 95

Ala Ala Glu Ile Tyr Gly Thr Lys Glu Ser Pro Gln Thr His Tyr Tyr

100 105 110

Ala Val Ala Val Val Lys Lys Gly Ser Asn Phe Gln Leu Asp Gln Leu

115 120 125

Gln Gly Arg Lys Ser Cys His Thr Gly Leu Gly Arg Ser Ala Gly Trp

130 135 140

Val Ile Pro Met Gly Ile Leu Arg Pro Tyr Leu Ser Trp Thr Glu Ser

145 150 155 160

Leu Glu Pro Leu Gln Gly Ala Val Ala Lys Phe Phe Ser Ala Ser Cys

165 170 175

Val Pro Cys Ile Asp Arg Gln Ala Tyr Pro Asn Leu Cys Gln Leu Cys

180 185 190

Lys Gly Glu Gly Glu Asn Gln Cys Ala Cys Ser Ser Arg Glu Pro Tyr

195 200 205

Phe Gly Tyr Ser Gly Ala Phe Lys Cys Leu Gln Asp Gly Ala Gly Asp

210 215 220

Val Ala Phe Val Lys Glu Thr Thr Val Phe Glu Asn Leu Pro Glu Lys

225 230 235 240

Ala Asp Arg Asp Gln Tyr Glu Leu Leu Cys Leu Asn Asn Ser Arg Ala

245 250 255

Pro Val Asp Ala Phe Lys Glu Cys His Leu Ala Gln Val Pro Ser His

260 265 270

Ala Val Val Ala Arg Ser Val Asp Gly Lys Glu Asp Leu Ile Trp Lys

275 280 285

Leu Leu Ser Lys Ala Gln Glu Lys Phe Gly Lys Asn Lys Ser Arg Ser

290 295 300

Phe Gln Leu Phe Gly Ser Pro Pro Gly Gln Arg Asp Leu Leu Phe Lys

305 310 315 320

Asp Ser Ala Leu Gly Phe Leu Arg Ile Pro Ser Lys Val Asp Ser Ala

325 330 335

Leu Tyr Leu Gly Ser Arg Tyr Leu Thr Thr Leu Lys Asn Leu Arg Glu

340 345 350

Thr Ala Glu Glu Val Lys Ala Arg Tyr Thr Arg Val Val Trp Cys Ala

355 360 365

Val Gly Pro Glu Glu Gln Lys Lys Cys Gln Gln Trp Ser Gln Gln Ser

370 375 380

Gly Gln Asn Val Thr Cys Ala Thr Ala Ser Thr Thr Asp Asp Cys Ile

385 390 395 400

Val Leu Val Leu Lys Gly Glu Ala Asp Ala Leu Asn Leu Asp Gly Gly

405 410 415

Tyr Ile Tyr Thr Ala Gly Lys Cys Gly Leu Val Pro Val Leu Ala Glu

420 425 430

Asn Arg Lys Thr Ser Lys Tyr Ser Ser Leu Asp Cys Val Leu Arg Pro

435 440 445

Thr Glu Gly Tyr Leu Ala Val Ala Val Val Lys Lys Ala Asn Glu Gly

450 455 460

Leu Thr Trp Asn Ser Leu Lys Asp Lys Lys Ser Cys His Thr Ala Val

465 470 475 480

Asp Arg Thr Ala Gly Trp Asn Ile Pro Met Gly Leu Ile Val Asn Gln

485 490 495

Thr Gly Ser Cys Ala Phe Asp Glu Phe Phe Ser Gln Ser Cys Ala Pro

500 505 510

Gly Arg Asp Pro Lys Ser Arg Leu Cys Ala Leu Cys Ala Gly Asp Asp

515 520 525

Gln Gly Leu Asp Lys Cys Val Pro Asn Ser Lys Glu Lys Tyr Tyr Gly

530 535 540

Tyr Thr Gly Ala Phe Arg Cys Leu Ala Glu Asp Val Gly Asp Val Ala

545 550 555 560

Phe Val Lys Asn Asp Thr Val Trp Glu Asn Thr Asn Gly Glu Ser Thr

565 570 575

Ala Asp Trp Ala Lys Asn Leu Asn Arg Glu Asp Phe Arg Leu Leu Cys

580 585 590

Leu Asp Gly Thr Arg Lys Pro Val Thr Glu Ala Gln Ser Cys His Leu

595 600 605

Ala Val Ala Pro Asn His Ala Val Val Ser Arg Ser Asp Arg Ala Ala

610 615 620

His Val Lys Gln Val Leu Leu His Gln Gln Ala Leu Phe Gly Lys Asn

625 630 635 640

Gly Lys Asn Cys Pro Asp Lys Phe Cys Leu Phe Lys Ser Glu Thr Lys

645 650 655

Asn Leu Leu Phe Asn Asp Asn Thr Glu Cys Leu Ala Lys Leu Gly Gly

660 665 670

Arg Pro Thr Tyr Glu Glu Tyr Leu Gly Thr Glu Tyr Val Thr Ala Ile

675 680 685

Ala Asn Leu Lys Lys Cys Ser Thr Ser Pro Leu Leu Glu Ala Cys Ala

690 695 700

Phe Leu Thr Arg

705

<210> 109

<211> 2357

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Lactoferrin cDNA

<400> 109

gcagagcctt cgttccggag tcgccccagg accccagcca tgaagctctt cgtccccgcc 60

ctgctgtccc ttggagccct tggactgtgt ctggctgccc cgaggaaaaa cgttcgatgg 120

tgtaccatct cccaacccga gtggttcaaa tgccgccgat ggcagtggag gatgaagaag 180

ctgggtgctc cctctatcac ctgtgtgagg agggcctttg ccttggaatg tatccgggcc 240

atcgcggaga aaaaggcgga tgctgtgacc ctggatggtg gcatggtgtt tgaggcgggc 300

cgggacccct acaaactgcg gccagtagca gcagagatct atgggacgaa agagtctccc 360

caaacccact attatgctgt ggccgtcgtg aagaagggca gcaactttca gctggaccag 420

ctgcaaggcc ggaagtcctg ccatacgggc cttggcaggt ccgctgggtg ggtcatccct 480

atgggaatcc ttcgcccgta cttgagctgg acagagtcac tcgagcccct ccagggagct 540

gtggctaaat tcttctctgc cagctgtgtt ccctgcattg atagacaagc ataccccaac 600

ctgtgtcaac tgtgcaaggg ggagggggag aaccagtgtg cctgctcctc ccgggaacca 660

tacttcggtt attctggtgc cttcaagtgt ctgcaggacg gggctggaga cgtggctttt 720

gttaaagaga cgacagtgtt tgagaacttg ccagagaagg ctgacaggga ccagtatgag 780

cttctctgcc tgaacaacag tcgggcgcca gtggatgcgt tcaaggagtg ccacctggcc 840

caggtccctt ctcatgctgt cgtggcccga agtgtggatg gcaaggaaga cttgatctgg 900

aagcttctca gcaaggcgca ggagaaattt ggaaaaaaca agtctcggag cttccagctc 960

tttggctctc cacccggcca gagggacctg ctgttcaaag actctgctct tgggtttttg 1020

aggatcccct cgaaggtaga ttcggcgctg taccttggct cccgctactt gaccaccttg 1080

aagaacctca gggaaactgc ggaggaggtg aaggcgcggt acaccagggt cgtgtggtgt 1140

gccgtgggac ccgaggagca gaagaagtgc cagcagtgga gccagcagag cggccagaac 1200

gtgacctgtg ccacggcgtc caccaccgac gactgcatcg tcctggtgct gaaaggggaa 1260

gcagatgccc tgaacttgga tggaggatat atctacactg cgggcaagtg tggcctggtg 1320

cctgtcctgg cagagaaccg gaaaacctcc aaatacagta gcctagattg tgtgctgaga 1380

ccaacagaag ggtaccttgc cgtggcagtt gtcaagaaag caaatgaggg gctcacatgg 1440

aattctctga aagacaagaa gtcgtgccac accgccgtgg acaggactgc aggctggaac 1500

atccccatgg gcctgatcgt caaccagaca ggctcctgcg catttgatga attctttagt 1560

cagagctgtg cccctgggcg tgacccgaaa tccagactct gtgccttgtg tgctggcgat 1620

gaccagggcc tggacaagtg tgtgcccaac tctaaggaga agtactatgg ctataccggg 1680

gctttcaggt gcctggctga ggacgttggg gacgttgcct ttgtgaaaaa cgacacagtc 1740

tgggagaaca cgaatggaga gagcactgca gactgggcta agaacttgaa tcgtgaggac 1800

ttcaggttgc tctgcctcga tggcaccagg aagcctgtga cggaggctca gagctgccac 1860

ctggcggtgg ccccgaatca cgctgtggtg tctcggagcg atagggcagc acacgtgaaa 1920

caggtgctgc tccaccagca ggctctgttt gggaaaaatg gaaaaaactg cccggacaag 1980

ttttgtttgt tcaaatctga aaccaaaaac cttctgttca atgacaacac tgagtgtctg 2040

gccaaacttg gaggcagacc aacgtatgaa gaatatttgg ggacagagta tgtcacggcc 2100

attgccaacc tgaaaaaatg ctcaacctcc ccgcttctgg aagcctgcgc cttcctgacg 2160

aggtaaagcc tgcaaagaag ctagcctgcc tccctgggcc tcagctcctc cctgctctca 2220

gccccaatct ccaggcgcga gggaccttcc tctcccttcc tgaagtcgga tttttgccaa 2280

gctcatcagt atttacaatt ccctgctgtc attttagcaa gaaataaaat tagaaatgct 2340

gttgattttc attccct 2357

<210> 110

<211> 708

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> lactoferrin

<400> 110

Met Lys Leu Phe Val Pro Ala Leu Leu Ser Leu Gly Ala Leu Gly Leu

1 5 10 15

Cys Leu Ala Ala Pro Arg Lys Asn Val Arg Trp Cys Ala Ile Ser Leu

20 25 30

Pro Glu Trp Ser Lys Cys Tyr Gln Trp Gln Arg Arg Met Arg Lys Leu

35 40 45

Gly Ala Pro Ser Ile Thr Cys Val Arg Arg Thr Ser Ala Leu Glu Cys

50 55 60

Ile Arg Ala Ile Ala Gly Lys Asn Ala Asp Ala Val Thr Leu Asp Ser

65 70 75 80

Gly Met Val Phe Glu Ala Gly Arg Asp Pro Tyr Lys Leu Arg Pro Val

85 90 95

Ala Ala Glu Ile Tyr Gly Thr Glu Lys Ser Pro Gln Thr His Tyr Tyr

100 105 110

Ala Val Ala Val Val Lys Lys Gly Ser Asn Phe Lys Leu Asp Gln Leu

115 120 125

Gln Gly Gln Lys Ser Cys His Met Gly Leu Gly Arg Ser Ala Gly Trp

130 135 140

Asn Ile Pro Val Gly Ile Leu Arg Pro Pro Leu Ser Trp Thr Glu Ser

145 150 155 160

Ala Glu Pro Leu Gln Gly Ala Val Ala Arg Phe Phe Ser Ala Ser Cys

165 170 175

Val Pro Cys Val Asp Gly Lys Ala Tyr Pro Asn Leu Cys Gln Leu Cys

180 185 190

Lys Gly Val Gly Glu Asn Lys Cys Ala Cys Ser Ser Gln Glu Pro Tyr

195 200 205

Phe Gly Tyr Ser Gly Ala Phe Lys Cys Leu Gln Asp Gly Ala Gly Asp

210 215 220

Val Ala Phe Val Lys Glu Thr Thr Val Phe Glu Asn Leu Pro Glu Lys

225 230 235 240

Ala Asp Arg Asp Gln Tyr Glu Leu Leu Cys Leu Asn Asn Thr Arg Ala

245 250 255

Pro Val Asp Ala Phe Lys Glu Cys His Leu Ala Gln Val Pro Ser His

260 265 270

Ala Val Val Ala Arg Ser Val Asp Gly Lys Glu Asn Leu Ile Trp Glu

275 280 285

Leu Leu Arg Lys Ala Gln Glu Lys Phe Gly Lys Asn Lys Ser Gln Arg

290 295 300

Phe Gln Leu Phe Gly Ser Pro Glu Gly Arg Arg Asp Leu Leu Phe Lys

305 310 315 320

Asp Ser Ala Leu Gly Phe Val Arg Ile Pro Ser Lys Val Asp Ser Ala

325 330 335

Leu Tyr Leu Gly Ser Arg Tyr Leu Thr Ala Leu Lys Asn Leu Arg Glu

340 345 350

Thr Ala Glu Glu Leu Lys Ala Arg Cys Thr Arg Val Val Trp Cys Ala

355 360 365

Val Gly Pro Glu Glu Gln Ser Lys Cys Gln Gln Trp Ser Glu Gln Ser

370 375 380

Gly Gln Asn Val Thr Cys Ala Thr Ala Ser Thr Thr Asp Asp Cys Ile

385 390 395 400

Ala Leu Val Leu Lys Gly Glu Ala Asp Ala Leu Ser Leu Gly Gly Gly

405 410 415

Tyr Ile Tyr Thr Ala Gly Lys Cys Gly Leu Val Pro Val Met Ala Glu

420 425 430

Asn Arg Lys Ser Ser Lys Tyr Ser Ser Leu Asp Cys Val Leu Arg Pro

435 440 445

Thr Glu Gly Tyr Leu Ala Val Ala Val Val Lys Lys Ala Asn Glu Gly

450 455 460

Leu Thr Trp Asn Ser Leu Lys Gly Lys Lys Ser Cys His Thr Ala Val

465 470 475 480

Asp Arg Thr Ala Gly Trp Asn Ile Pro Met Gly Leu Ile Ala Asn Gln

485 490 495

Thr Gly Ser Cys Ala Phe Asp Glu Phe Phe Ser Gln Ser Cys Ala Pro

500 505 510

Gly Ala Asp Pro Lys Ser Ser Leu Cys Ala Leu Cys Ala Gly Asp Asp

515 520 525

Gln Gly Leu Asp Lys Cys Val Pro Asn Ser Lys Glu Lys Tyr Tyr Gly

530 535 540

Tyr Thr Gly Ala Phe Arg Cys Leu Ala Glu Asp Val Gly Asp Val Ala

545 550 555 560

Phe Val Lys Asn Asp Thr Val Trp Glu Asn Thr Asn Gly Glu Ser Ser

565 570 575

Ala Asp Trp Ala Lys Asn Leu Asn Arg Glu Asp Phe Arg Leu Leu Cys

580 585 590

Leu Asp Gly Thr Thr Lys Pro Val Thr Glu Ala Gln Ser Cys Tyr Leu

595 600 605

Ala Val Ala Pro Asn His Ala Val Val Ser Arg Ser Asp Arg Ala Ala

610 615 620

His Val Glu Gln Val Leu Leu His Gln Gln Ala Leu Phe Gly Lys Asn

625 630 635 640

Gly Lys Asn Cys Pro Asp Gln Phe Cys Leu Phe Lys Ser Glu Thr Lys

645 650 655

Asn Leu Leu Phe Asn Asp Asn Thr Glu Cys Leu Ala Lys Leu Gly Gly

660 665 670

Arg Pro Thr Tyr Glu Lys Tyr Leu Gly Thr Glu Tyr Val Thr Ala Ile

675 680 685

Ala Asn Leu Lys Lys Cys Ser Thr Ser Pro Leu Leu Glu Ala Cys Ala

690 695 700

Phe Leu Thr Arg

705

<210> 111

<211> 2411

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> lactoferrin

<400> 111

agcactggat aaagggacgc agaacgaggg caggtggcag agcttcgttc cggagtcgcc 60

ccaggacccc agacatgaag ctcttcgtcc ccgccctgct gtcccttgga gcccttggac 120

tgtgtctggc tgccccgagg aaaaacgttc gatggtgtgc catctcactg ccggagtggt 180

ccaaatgcta ccaatggcag aggaggatga ggaagctggg tgctccctct atcacctgtg 240

tgaggaggac ctctgccttg gaatgtatcc gggccatcgc gggaaaaaat gcggatgctg 300

tgaccctgga tagtggcatg gtgtttgagg cgggccggga cccctacaaa ctgcggccag 360

tagcggcaga gatctatggg acagaaaaat ctccccaaac ccactattat gctgtggccg 420

tcgtgaagaa gggcagcaac tttaagctgg accagctgca aggtcagaag tcctgccaca 480

tgggccttgg caggtccgct gggtggaaca tccctgtggg aatccttcgc ccgcccttga 540

gctggacaga gtcggccgag cccctccagg gagctgtggc tagattcttc tctgccagct 600

gtgttccctg cgttgatgga aaagcgtacc ccaacctgtg tcaactgtgc aagggggtgg 660

gagagaacaa gtgtgcctgc tcctcccagg aaccatactt tggttattct ggtgccttca 720

agtgcctgca ggacggggct ggagacgtgg cttttgtcaa ggagacgaca gtgtttgaga 780

acttgccaga gaaggctgac agggaccagt atgagcttct ctgcctaaac aacactcggg 840

cgccagtgga tgcattcaag gagtgccacc tggcccaggt cccttctcat gctgttgtgg 900

cccgcagtgt ggatggcaag gagaacttga tctgggagct tctcaggaag gcacaggaga 960

agtttggaaa aaacaagtct cagcgcttcc agctctttgg ctctccagaa ggccggaggg 1020

acctgctatt caaagactct gcccttgggt ttgtgaggat cccctcaaaa gtagattcgg 1080

cgctgtacct gggctcccgt tacttgaccg ccttgaagaa cctcagggaa accgcggagg 1140

agttgaaggc gcggtgcacg cgggtcgtgt ggtgcgcggt gggacccgag gagcagagta 1200

agtgccagca gtggagcgag cagagcggcc agaacgtgac ctgtgccacg gcctccacca 1260

ccgacgactg catcgccctg gtgctgaaag gggaagcgga cgccctgagc ttgggtggag 1320

gatatatcta cactgccggc aagtgcggcc tggtgcctgt catggcagag aaccggaaat 1380

cctccaaata cagtagccta gattgtgtgc tgagaccaac ggaagggtac cttgccgtgg 1440

cagttgtcaa gaaagcaaat gaggggctca cttggaattc tctgaaaggc aagaagtcgt 1500

gccacaccgc cgtggacagg actgcaggct ggaacatccc catgggcctg atcgccaacc 1560

agacaggctc ctgcgcattt gatgaattct ttagtcagag ctgtgcccct ggggccgacc 1620

cgaaatccag cctctgtgca ttgtgtgccg gcgatgacca gggcctggac aagtgtgtgc 1680

ccaactctaa ggagaagtac tatggctaca ccggggcttt caggtgcctg gctgaggacg 1740

ttggggacgt tgcatttgtg aaaaacgaca cagtctggga gaacacaaat ggagagagct 1800

ctgcagactg ggctaagaac ttgaatcgcg aggacttcag gctgctctgc ctcgatggca 1860

ccacgaagcc tgtgacggag gctcagagct gctacctggc ggtggccccg aatcacgctg 1920

tggtgtctcg gagcgatagg gcagcgcacg tggaacaggt gctgctccac cagcaggctc 1980

tgtttgggaa aaatggaaaa aactgcccgg accagttttg tttgttcaaa tctgaaacca 2040

aaaaccttct gttcaatgac aacactgagt gtctggccaa acttggaggc agaccaacgt 2100

atgaaaaata tttggggaca gagtatgtca cggccattgc caacctgaaa aaatgctcaa 2160

cctccccgct tctggaagcc tgcgccttcc tgacgaggta aagcctgcaa agaagctagc 2220

ctgcctcccc gggcctcagc tcctccctgc tctcagcccc agtcttcagg cgcgagggac 2280

cttcctctcc cttcctgaag tcggattttt gccaagctca tcagtattca caattccctg 2340

ctgtcatctt agcaagaaat taaattagaa atgctgttga ttttcattcc ctaaaaaaaa 2400

aaaaaaaaaa a 2411

<210> 112

<211> 711

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> lactoferrin

<400> 112

Met Lys Leu Val Phe Leu Val Leu Leu Phe Leu Gly Ala Leu Gly Leu

1 5 10 15

Cys Leu Ala Gly Arg Arg Arg Arg Ser Val Gln Trp Cys Thr Val Ser

20 25 30

Gln Pro Glu Ala Thr Lys Cys Phe Gln Trp Gln Arg Asn Met Arg Arg

35 40 45

Val Arg Gly Pro Pro Val Ser Cys Ile Lys Arg Asp Ser Pro Ile Gln

50 55 60

Cys Ile Gln Ala Ile Ala Glu Asn Arg Ala Asp Ala Val Thr Leu Asp

65 70 75 80

Gly Gly Phe Ile Tyr Glu Ala Gly Leu Ala Pro Tyr Lys Leu Arg Pro

85 90 95

Val Ala Ala Glu Val Tyr Gly Thr Glu Arg Gln Pro Arg Thr His Tyr

100 105 110

Tyr Ala Val Ala Val Val Lys Lys Gly Gly Ser Phe Gln Leu Asn Glu

115 120 125

Leu Gln Gly Leu Lys Ser Cys His Thr Gly Leu Arg Arg Thr Ala Gly

130 135 140

Trp Asn Val Pro Ile Gly Thr Leu Arg Pro Phe Leu Asn Trp Thr Gly

145 150 155 160

Pro Pro Glu Pro Ile Glu Ala Ala Val Ala Arg Phe Phe Ser Ala Ser

165 170 175

Cys Val Pro Gly Ala Asp Lys Gly Gln Phe Pro Asn Leu Cys Arg Leu

180 185 190

Cys Ala Gly Thr Gly Glu Asn Lys Cys Ala Phe Ser Ser Gln Glu Pro

195 200 205

Tyr Phe Ser Tyr Ser Gly Ala Phe Lys Cys Leu Arg Asp Gly Ala Gly

210 215 220

Asp Val Ala Phe Ile Arg Glu Ser Thr Val Phe Glu Asp Leu Ser Asp

225 230 235 240

Glu Ala Glu Arg Asp Glu Tyr Glu Leu Leu Cys Pro Asp Asn Thr Arg

245 250 255

Lys Pro Val Asp Lys Phe Lys Asp Cys His Leu Ala Arg Val Pro Ser

260 265 270

His Ala Val Val Ala Arg Ser Val Asn Gly Lys Glu Asp Ala Ile Trp

275 280 285

Asn Leu Leu Arg Gln Ala Gln Glu Lys Phe Gly Lys Asp Lys Ser Pro

290 295 300

Lys Phe Gln Leu Phe Gly Ser Pro Ser Gly Gln Lys Asp Leu Leu Phe

305 310 315 320

Lys Asp Ser Ala Ile Gly Phe Ser Arg Val Pro Pro Arg Ile Asp Ser

325 330 335

Gly Leu Tyr Leu Gly Ser Gly Tyr Phe Thr Ala Ile Gln Asn Leu Arg

340 345 350

Lys Ser Glu Glu Glu Val Ala Ala Arg Arg Ala Arg Val Val Trp Cys

355 360 365

Ala Val Gly Glu Gln Glu Leu Arg Lys Cys Asn Gln Trp Ser Gly Leu

370 375 380

Ser Glu Gly Ser Val Thr Cys Ser Ser Ala Ser Thr Thr Glu Asp Cys

385 390 395 400

Ile Ala Leu Val Leu Lys Gly Glu Ala Asp Ala Met Ser Leu Asp Gly

405 410 415

Gly Tyr Val Tyr Thr Ala Gly Lys Cys Gly Leu Val Pro Val Leu Ala

420 425 430

Glu Asn Tyr Lys Ser Gln Gln Ser Ser Asp Pro Asp Pro Asn Cys Val

435 440 445

Asp Arg Pro Val Glu Gly Tyr Leu Ala Val Ala Val Val Arg Arg Ser

450 455 460

Asp Thr Ser Leu Thr Trp Asn Ser Val Lys Gly Lys Lys Ser Cys His

465 470 475 480

Thr Ala Val Asp Arg Thr Ala Gly Trp Asn Ile Pro Met Gly Leu Leu

485 490 495

Phe Asn Gln Thr Gly Ser Cys Lys Phe Asp Glu Tyr Phe Ser Gln Ser

500 505 510

Cys Ala Pro Gly Ser Asp Pro Arg Ser Asn Leu Cys Ala Leu Cys Ile

515 520 525

Gly Asp Glu Gln Gly Glu Asn Lys Cys Val Pro Asn Ser Asn Glu Arg

530 535 540

Tyr Tyr Gly Tyr Thr Gly Ala Phe Arg Cys Leu Ala Glu Asn Ala Gly

545 550 555 560

Asp Val Ala Phe Val Lys Asp Val Thr Val Leu Gln Asn Thr Asp Gly

565 570 575

Asn Asn Asn Asp Ala Trp Ala Lys Asp Leu Lys Leu Ala Asp Phe Ala

580 585 590

Leu Leu Cys Leu Asp Gly Lys Arg Lys Pro Val Thr Glu Ala Arg Ser

595 600 605

Cys His Leu Ala Met Ala Pro Asn His Ala Val Val Ser Arg Met Asp

610 615 620

Lys Val Glu Arg Leu Lys Gln Val Leu Leu His Gln Gln Ala Lys Phe

625 630 635 640

Gly Arg Asn Gly Ser Asp Cys Pro Asp Lys Phe Cys Leu Phe Gln Ser

645 650 655

Glu Thr Lys Asn Leu Leu Phe Asn Asp Asn Thr Glu Cys Leu Ala Arg

660 665 670

Leu His Gly Lys Thr Thr Tyr Glu Lys Tyr Leu Gly Pro Gln Tyr Val

675 680 685

Ala Gly Ile Thr Asn Leu Lys Lys Cys Ser Thr Ser Pro Leu Leu Glu

690 695 700

Ala Cys Glu Phe Leu Arg Lys

705 710

<210> 113

<211> 2259

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> Lactoferrin cDNA

<400> 113

cagaccgcag acatgaaact tgtcttcctc gtcctgctgt tcctcggggc cctcggactg 60

tgtctggctg gccgtaggag aaggagtgtt cagtggtgca ccgtatccca acccgaggcc 120

acaaaatgct tccaatggca aaggaatatg agaagagtgc gtggccctcc tgtcagctgc 180

ataaagagag actcccccat ccagtgtatc caggccattg cggaaaacag ggccgatgct 240

gtgacccttg atggtggttt catatacgag gcaggcctgg ccccctacaa actgcgacct 300

gtagcggcgg aagtctacgg gaccgaaaga cagccacgaa ctcactatta tgccgtggct 360

gtggtgaaga agggcggcag ctttcagctg aacgaactgc aaggtctgaa gtcctgccac 420

acaggccttc gcaggaccgc tggatggaat gtccctatag ggacacttcg tccattcttg 480

aattggacgg gtccacctga gcccattgag gcagctgtgg ccaggttctt ctcagccagc 540

tgtgttcccg gtgcagataa aggacagttc cccaacctgt gtcgcctgtg tgcggggaca 600

ggggaaaaca aatgtgcctt ctcctcccag gaaccgtact tcagctactc tggtgccttc 660

aagtgtctga gagacggggc tggagacgtg gcttttatca gagagagcac agtgtttgag 720

gacctgtcag acgaggctga aagggacgag tatgagttac tctgcccaga caacactcgg 780

aagccagtgg acaagttcaa agactgccat ctggcccggg tcccttctca tgccgttgtg 840

gcacgaagtg tgaatggcaa ggaggatgcc atctggaatc ttctccgcca ggcacaggaa 900

aagtttggaa aggacaagtc accgaaattc cagctctttg gctcccctag tgggcagaaa 960

gatctgctgt tcaaggactc tgccattggg ttttcgaggg tgcccccgag gatagattct 1020

gggctgtacc ttggctccgg ctacttcact gccatccaga acttgaggaa aagtgaggag 1080

gaagtggctg cccggcgtgc gcgggtcgtg tggtgtgcgg tgggcgagca ggagctgcgc 1140

aagtgtaacc agtggagtgg cttgagcgaa ggcagcgtga cctgctcctc ggcctccacc 1200

acagaggact gcatcgccct ggtgctgaaa ggagaagctg atgccatgag tttggatgga 1260

ggatatgtgt acactgcagg caaatgtggt ttggtgcctg tcctggcaga gaactacaaa 1320

tcccaacaaa gcagtgaccc tgatcctaac tgtgtggata gacctgtgga aggatatctt 1380

gctgtggcgg tggttaggag atcagacact agccttacct ggaactctgt gaaaggcaag 1440

aagtcctgcc acaccgccgt ggacaggact gcaggctgga atatccccat gggcctgctc 1500

ttcaaccaga cgggctcctg caaatttgat gaatatttca gtcaaagctg tgcccctggg 1560

tctgacccga gatctaatct ctgtgctctg tgtattggcg acgagcaggg tgagaataag 1620

tgcgtgccca acagcaatga gagatactac ggctacactg gggctttccg gtgcctggct 1680

gagaatgctg gagacgttgc atttgtgaaa gatgtcactg tcttgcagaa cactgatgga 1740

aataacaatg acgcatgggc taaggatttg aagctggcag actttgcgct gctgtgcctc 1800

gatggcaaac ggaagcctgt gactgaggct agaagctgcc atcttgccat ggccccgaat 1860

catgccgtgg tgtctcggat ggataaggtg gaacgcctga aacaggtgtt gctccaccaa 1920

caggctaaat ttgggagaaa tggatctgac tgcccggaca agttttgctt attccagtct 1980

gaaaccaaaa accttctgtt caatgacaac actgagtgtc tggccagact ccatggcaaa 2040

acaacatatg aaaaatattt gggaccacag tatgtcgcag gcattactaa tctgaaaaag 2100

tgctcaacct cccccctcct ggaagcctgt gaattcctca ggaagtaaaa ccgaagaaga 2160

tggcccagct ccccaagaaa gcctcagcca ttcactgccc ccagctcttc tccccaggtg 2220

tgttggggcc ttggctcccc tgctgaaggt ggggattgc 2259

<210> 114

<211> 708

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> lactoferrin

<400> 114

Met Lys Leu Phe Val Pro Ala Leu Leu Ser Leu Gly Ala Leu Gly Leu

1 5 10 15

Cys Leu Ala Ala Pro Arg Lys Asn Val Arg Trp Cys Ala Ile Ser Pro

20 25 30

Pro Glu Gly Ser Lys Cys Tyr Gln Trp Gln Arg Arg Met Arg Lys Leu

35 40 45

Gly Ala Pro Ser Ile Thr Cys Val Arg Arg Thr Ser Ala Leu Glu Cys

50 55 60

Ile Arg Ala Ile Ala Gly Lys Lys Ala Asp Ala Val Thr Leu Asp Ser

65 70 75 80

Gly Met Val Phe Glu Ala Gly Leu Asp Pro Tyr Lys Leu Arg Pro Val

85 90 95

Ala Ala Glu Ile Tyr Gly Thr Glu Lys Ser Pro Gln Thr His Tyr Tyr

100 105 110

Ala Val Ala Val Val Lys Lys Gly Ser Asn Phe Gln Leu Asp Gln Leu

115 120 125

Gln Gly Gln Lys Ser Cys His Met Gly Leu Gly Arg Ser Ala Gly Trp

130 135 140

Asn Ile Pro Met Gly Ile Leu Arg Pro Phe Leu Ser Trp Thr Glu Ser

145 150 155 160

Ala Glu Pro Leu Gln Gly Ala Val Ala Arg Phe Phe Ser Ala Ser Cys

165 170 175

Val Pro Cys Val Asp Gly Lys Ala Tyr Pro Asn Leu Cys Gln Leu Cys

180 185 190

Lys Gly Val Gly Glu Asn Lys Cys Ala Cys Ser Ser Gln Glu Pro Tyr

195 200 205

Phe Gly Tyr Ser Gly Ala Phe Lys Cys Leu Gln Asp Gly Ala Gly Asp

210 215 220

Val Ala Phe Val Lys Glu Thr Thr Val Phe Glu Asn Leu Pro Glu Lys

225 230 235 240

Ala Asp Arg Asp Gln Tyr Glu Leu Leu Cys Leu Asn Asn Thr Arg Ala

245 250 255

Pro Val Asp Ala Phe Lys Glu Cys His Leu Ala Gln Val Pro Ser His

260 265 270

Ala Val Val Ala Arg Ser Val Asp Gly Lys Glu Asn Leu Ile Trp Glu

275 280 285

Leu Leu Arg Lys Ala Gln Glu Lys Phe Gly Lys Asn Lys Ser Gln Arg

290 295 300

Phe Gln Leu Phe Gly Ser Pro Gln Gly Gln Lys Asp Leu Leu Phe Lys

305 310 315 320

Asp Ser Ala Leu Gly Phe Val Arg Ile Pro Ser Lys Val Asp Ser Ala

325 330 335

Leu Tyr Leu Gly Ser Arg Tyr Leu Thr Ala Leu Lys Asn Leu Arg Glu

340 345 350

Thr Ala Glu Glu Val Lys Ala Arg Cys Thr Arg Val Val Trp Cys Ala

355 360 365

Val Gly Pro Glu Glu His Ser Lys Cys Gln Gln Trp Ser Glu Gln Ser

370 375 380

Gly Gln Asn Val Thr Cys Ala Met Ala Ser Thr Thr Asp Asp Cys Ile

385 390 395 400

Ala Leu Val Leu Lys Gly Glu Ala Asp Ala Leu Ser Leu Asp Gly Gly

405 410 415

Tyr Ile Tyr Thr Ala Gly Lys Cys Gly Leu Val Pro Val Met Ala Glu

420 425 430

Asn Arg Glu Ser Ser Lys Tyr Ser Ser Leu Asp Cys Val Leu Arg Pro

435 440 445

Thr Glu Gly Tyr Leu Ala Val Ala Val Val Lys Lys Ala Asn Glu Gly

450 455 460

Leu Thr Trp Asn Ser Leu Lys Gly Lys Lys Ser Cys His Thr Ala Val

465 470 475 480

Asp Arg Thr Ala Gly Trp Asn Ile Pro Met Gly Leu Ile Ala Asn Gln

485 490 495

Thr Gly Ser Cys Ala Phe Asp Glu Phe Phe Ser Gln Ser Cys Ala Pro

500 505 510

Gly Ala Asp Pro Lys Ser Ser Leu Cys Ala Leu Cys Ala Gly Asp Asp

515 520 525

Gln Gly Leu Asn Lys Cys Val Pro Asn Ser Lys Glu Lys Tyr Tyr Gly

530 535 540

Tyr Thr Gly Ala Phe Arg Cys Leu Ala Glu Asp Val Gly Asp Val Ala

545 550 555 560

Phe Val Lys Asn Asp Thr Val Trp Glu Asn Thr Asn Gly Glu Ser Ser

565 570 575

Ala Asp Trp Ala Lys Asn Leu Asn Arg Glu Asp Phe Arg Leu Leu Cys

580 585 590

Leu Asp Gly Thr Thr Lys Pro Val Thr Glu Ala Gln Ser Cys Tyr Leu

595 600 605

Ala Val Ala Pro Asn His Ala Val Val Ser Arg Ser Asp Arg Ala Ala

610 615 620

His Val Glu Gln Val Leu Leu His Gln Gln Ala Leu Phe Gly Lys Asn

625 630 635 640

Gly Lys Asn Cys Pro Asp Gln Phe Cys Leu Phe Lys Ser Glu Thr Lys

645 650 655

Asn Leu Leu Phe Asn Asp Asn Thr Glu Cys Leu Ala Lys Leu Gly Gly

660 665 670

Arg Pro Thr Tyr Glu Lys Tyr Leu Gly Thr Glu Tyr Val Thr Ala Ile

675 680 685

Ala Asn Leu Lys Lys Cys Ser Thr Ser Pro Leu Leu Glu Ala Cys Ala

690 695 700

Phe Leu Thr Arg

705

<210> 115

<211> 2127

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> Lactoferrin cDN

<400> 115

atgaagctct tcgtccccgc cctgctgtcc cttggagccc ttggactgtg tctggctgcc 60

ccgaggaaaa acgttcgatg gtgtgccatc tcaccgccgg aggggtccaa atgctaccaa 120

tggcagagga ggatgaggaa gctgggtgct ccctctatca cctgtgtgag gaggacctct 180

gccctggaat gtatccgggc catcgcggga aaaaaggcgg atgctgtgac cctggatagt 240

ggcatggtgt ttgaggcggg cctggacccc tacaaactgc ggccagtagc ggcagagatc 300

tatgggacag aaaagtctcc ccaaacccac tattatgctg tggccgtcgt gaagaagggc 360

agcaactttc agctggacca gctgcaaggc cagaagtcct gccacatggg ccttggtagg 420

tccgctgggt ggaacatccc tatgggaatc cttcgcccgt tcttgagctg gacagagtcg 480

gccgagcccc tccagggagc tgtggctaga ttcttctctg ccagctgtgt tccctgcgtt 540

gatggaaaag cgtatcccaa cctgtgtcaa ctgtgcaagg gggtgggaga gaacaagtgt 600

gcctgctcct cccaggaacc atactttggt tattctggtg ccttcaagtg cctgcaggac 660

ggggctggag acgtggcttt tgtcaaggag acgacagtgt ttgagaactt gccagagaag 720

gctgacaggg accagtatga gcttctctgc ctaaacaaca ctcgggcgcc agtggatgca 780

ttcaaggagt gccacctggc ccaggtccct tctcatgctg ttgtggcccg cagtgtggat 840

ggcaaggaga acttgatctg ggagcttctc aggaaggcac aggagaagtt tggaaaaaac 900

aagtctcagc gcttccagct ctttggctct ccacaaggcc agaaggacct gctattcaaa 960

gactctgccc ttgggtttgt gaggatcccc tcaaaagtag attcggcgct gtacttaggc 1020

tcccgctact tgaccgcctt gaagaacctc agggaaaccg cggaagaagt gaaggcgcgg 1080

tgcactccgg tcgtgtggtg cgcggtggga ccccaggagc atagtaagtg ccagcagtgg 1140

agcgagcaga gcggccagaa cgtgacttgt gccatggcct ccaccaccga cgaatgcatc 1200

gccctggtgt tgaaagggga agcggacgcc ctgagcttgg atggaggata tatctacact 1260

gccggcaagt gtggcctggt gcctgtcatg gcagagaacc gggaatcctc caaatacagt 1320

agcctagatt gtgtgctgag accaacggaa gggtaccttg ccgtggcagt tgtcaagaaa 1380

gcaaatgagg ggctcacttg gaattctctg aaaggcaaga agtcgtgcca caccgccgtg 1440

gacaggactg caggctggaa catccccatg ggcctgatcg ccaaccagac aggctcctgc 1500

gcatttgatg aattctttag tcagagctgt gcccctgggg ccgacccgaa atccagcctc 1560

tgtgcattgt gtgccggcga tgaccagggc ctgaacaagt gtgtgcccaa ctctaaggag 1620

aagtactatg gctacaccgg ggctttcagg tgcctggctg aggacgttgg ggacgttgca 1680

tttgtgaaaa acgacacagt ctgggagaac acgaatggag agagctctgc agactgggct 1740

aagaacttga atcgcgagga cttcaggttg ctctgcctcg atggcaccac gaagcctgtg 1800

acggaggctc agagctgcta cctggcggtg gccccgaatc acgctgtggt gtctcggagc 1860

gatagggcag cacacgtgga acaggtgctg ctccaccagc aggctctgtt cgggaaaaat 1920

ggaaaaaact gcccggacca gttttgtttg ttcaaatctg aaaccaaaaa ccttctgttc 1980

aatgacaaca ctgagtgtct ggccaaactt ggaggcagac caacgtatga aaaatatttg 2040

gggacagagt atgtcacggc cattgccaac ctgaaaaaat gctcaacctc cccgcttctg 2100

gaagcctgcg ccttcctgac gaggtaa 2127

<210> 116

<211> 704

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> transferrin

<400> 116

Met Arg Pro Ala Val Arg Ala Leu Leu Ala Cys Ala Val Leu Gly Leu

1 5 10 15

Cys Leu Ala Asp Pro Glu Arg Thr Val Arg Trp Cys Thr Ile Ser Thr

20 25 30

His Glu Ala Asn Lys Cys Ala Ser Phe Arg Glu Asn Val Leu Arg Ile

35 40 45

Leu Glu Ser Gly Pro Phe Val Ser Cys Val Lys Lys Thr Ser His Met

50 55 60

Asp Cys Ile Lys Ala Ile Ser Asn Asn Glu Ala Asp Ala Val Thr Leu

65 70 75 80

Asp Gly Gly Leu Val Tyr Glu Ala Gly Leu Lys Pro Asn Asn Leu Lys

85 90 95

Pro Val Val Ala Glu Phe His Gly Thr Lys Asp Asn Pro Gln Thr His

100 105 110

Tyr Tyr Ala Val Ala Val Val Lys Lys Asp Thr Asp Phe Lys Leu Asn

115 120 125

Glu Leu Arg Gly Lys Lys Ser Cys His Thr Gly Leu Gly Arg Ser Ala

130 135 140

Gly Trp Asn Ile Pro Met Ala Lys Leu Tyr Lys Glu Leu Pro Asp Pro

145 150 155 160

Gln Glu Ser Ile Gln Arg Ala Ala Ala Asn Phe Phe Ser Ala Ser Cys

165 170 175

Val Pro Cys Ala Asp Gln Ser Ser Phe Pro Lys Leu Cys Gln Leu Cys

180 185 190

Ala Gly Lys Gly Thr Asp Lys Cys Ala Cys Ser Asn His Glu Pro Tyr

195 200 205

Phe Gly Tyr Ser Gly Ala Phe Lys Cys Leu Met Glu Gly Ala Gly Asp

210 215 220

Val Ala Phe Val Lys His Ser Thr Val Phe Asp Asn Leu Pro Asn Pro

225 230 235 240

Glu Asp Arg Lys Asn Tyr Glu Leu Leu Cys Gly Asp Asn Thr Arg Lys

245 250 255

Ser Val Asp Asp Tyr Gln Glu Cys Tyr Leu Ala Met Val Pro Ser His

260 265 270

Ala Val Val Ala Arg Thr Val Gly Gly Lys Glu Asp Val Ile Trp Glu

275 280 285

Leu Leu Asn His Ala Gln Glu His Phe Gly Lys Asp Lys Pro Asp Asn

290 295 300

Phe Gln Leu Phe Gln Ser Pro His Gly Lys Asp Leu Leu Phe Lys Asp

305 310 315 320

Ser Ala Asp Gly Phe Leu Lys Ile Pro Ser Lys Met Asp Phe Glu Leu

325 330 335

Tyr Leu Gly Tyr Glu Tyr Val Thr Ala Leu Gln Asn Leu Arg Glu Ser

340 345 350

Lys Pro Pro Asp Ser Ser Lys Asp Glu Cys Met Val Lys Trp Cys Ala

355 360 365

Ile Gly His Gln Glu Arg Thr Lys Cys Asp Arg Trp Ser Gly Phe Ser

370 375 380

Gly Gly Ala Ile Glu Cys Glu Thr Ala Glu Asn Thr Glu Glu Cys Ile

385 390 395 400

Ala Lys Ile Met Lys Gly Glu Ala Asp Ala Met Ser Leu Asp Gly Gly

405 410 415

Tyr Leu Tyr Ile Ala Gly Lys Cys Gly Leu Val Pro Val Leu Ala Glu

420 425 430

Asn Tyr Lys Thr Glu Gly Glu Ser Cys Lys Asn Thr Pro Glu Lys Gly

435 440 445

Tyr Leu Ala Val Ala Val Val Lys Thr Ser Asp Ala Asn Ile Asn Trp

450 455 460

Asn Asn Leu Lys Asp Lys Lys Ser Cys His Thr Ala Val Asp Arg Thr

465 470 475 480

Ala Gly Trp Asn Ile Pro Met Gly Leu Leu Tyr Ser Lys Ile Asn Asn

485 490 495

Cys Lys Phe Asp Glu Phe Phe Ser Ala Gly Cys Ala Pro Gly Ser Pro

500 505 510

Arg Asn Ser Ser Leu Cys Ala Leu Cys Ile Gly Ser Glu Lys Gly Thr

515 520 525

Gly Lys Glu Cys Val Pro Asn Ser Asn Glu Arg Tyr Tyr Gly Tyr Thr

530 535 540

Gly Ala Phe Arg Cys Leu Val Glu Lys Gly Asp Val Ala Phe Val Lys

545 550 555 560

Asp Gln Thr Val Ile Gln Asn Thr Asp Gly Asn Asn Asn Glu Ala Trp

565 570 575

Ala Lys Asn Leu Lys Lys Glu Asn Phe Glu Val Leu Cys Lys Asp Gly

580 585 590

Thr Arg Lys Pro Val Thr Asp Ala Glu Asn Cys His Leu Ala Arg Gly

595 600 605

Pro Asn His Ala Val Val Ser Arg Lys Asp Lys Ala Thr Cys Val Glu

610 615 620

Lys Ile Leu Asn Lys Gln Gln Asp Asp Phe Gly Lys Ser Val Thr Asp

625 630 635 640

Cys Thr Ser Asn Phe Cys Leu Phe Gln Ser Asn Ser Lys Asp Leu Leu

645 650 655

Phe Arg Asp Asp Thr Lys Cys Leu Ala Ser Ile Ala Lys Lys Thr Tyr

660 665 670

Asp Ser Tyr Leu Gly Asp Asp Tyr Val Arg Ala Met Thr Asn Leu Arg

675 680 685

Gln Cys Ser Thr Ser Lys Leu Leu Glu Ala Cys Thr Phe His Lys Pro

690 695 700

<210> 117

<211> 2584

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> transferrin cDNA

<400> 117

gctcggccag ccccgcggga gatatagggg cgcggggagc agggaggcta cgcagaagcc 60

ggtcggtctg tgctctgcac tccacattac agaccgggga ggatgaggcc cgctgtccgc 120

gctctgttag cctgcgcggt tctggggctg tgtctggcgg accctgagag aactgtgaga 180

tggtgcacca tttcaactca tgaggccaat aagtgtgcca gttttcgtga aaatgtgctt 240

cgtatcttgg agagtggtcc ttttgtctcc tgtgtgaaga aaacctcaca catggattgc 300

atcaaggcta tctcgaataa cgaagcagat gctgtgacgt tggatggagg tttggtgtat 360

gaggcaggcc tgaagcccaa caacctgaag cctgtggtgg cagagttcca tgggacaaaa 420

gacaacccac aaactcacta ttatgctgtg gcggtggtga agaaggacac tgacttcaag 480

ctgaatgagc tcagaggcaa gaagtcctgc cacacaggcc tcggcaggtc cgctgggtgg 540

aacatcccca tgggcaaact ttataaggaa ttgcctgatc cacaggaatc aattcagaga 600

gctgcggcca acttcttctc tgccagctgt gtcccctgtg cggatcaatc atcatttccc 660

aaactctgtc aactgtgtgc ggggaaaggg acagacaagt gtgcctgctc caaccacgaa 720

ccatacttcg gctactcagg ggcctttaaa tgtctgatgg agggcgctgg ggatgtggcc 780

tttgtcaagc actcaacggt attcgacaac ctgcctaacc cagaagacag gaaaaactat 840

gagctgctct gcggggacaa cacccggaag tctgtagatg attaccaaga atgctacttg 900

gcaatggttc cttcccatgc ggttgtggct cgaactgtgg gcggcaagga ggatgtgatc 960

tgggaacttc tcaatcacgc ccaggaacat tttggcaaag ataaaccaga caatttccag 1020

cttttccaat cccctcatgg gaaggacctg ctgtttaagg actctgctga tgggttttta 1080

aagattcctt ctaagatgga ttttgagctg tacttgggat atgaatatgt cactgctctt 1140

cagaatctaa gagaaagtaa acccccggac tcctcaaagg atgagtgcat ggtgaagtgg 1200

tgtgcaattg gtcaccagga gaggacaaag tgtgatcggt ggagtgggtt cagcggcggg 1260

gcaatagagt gtgaaacagc agagaacact gaagagtgca tcgccaagat catgaaagga 1320

gaagctgatg ccatgagctt ggacggaggc tacctctaca tagcaggcaa atgtggcctg 1380

gtgcctgtcc tggcagagaa ctacaaaact gagggtgaaa gttgcaagaa cacaccagaa 1440

aaagggtatc ttgctgtagc tgtggttaaa acatcagatg ctaatatcaa ctggaacaat 1500

ctgaaagaca agaagtcctg ccacactgca gtagacagaa ccgctggctg gaacatcccc 1560

atgggtttgc tctacagcaa gattaataac tgtaaatttg atgaattttt cagcgcaggt 1620

tgtgcacctg gatctccgcg aaattccagt ctctgtgctc tgtgcattgg ctcagagaag 1680

ggtacaggaa aggagtgtgt tcccaacagc aatgaaagat actatggcta tacaggggct 1740

ttcaggtgtc tggtggagaa gggagacgtg gcctttgtga aggaccagac tgtcatacag 1800

aacactgacg gaaataataa tgaagcatgg gcaaaaaatc tgaagaagga aaattttgaa 1860

gtactatgca aagatggcac caggaaacct gtgacagatg ctgagaactg ccacctggcc 1920

cgaggcccga atcatgctgt ggtctcacgg aaagataagg caacttgtgt ggagaaaata 1980

ttaaacaaac agcaggatga ttttggaaaa tctgtaaccg actgcacgag caatttttgt 2040

ttattccaat caaattccaa ggaccttctg ttcagggatg acactaaatg tttggcttca 2100

attgcgaaaa aaacatatga ctcctactta ggggatgact acgtcagagc tatgaccaac 2160

ctgagacaat gctcaacctc aaaactcctg gaagcatgca ctttccacaa accttaaaat 2220

ccaagagtgg agccaacacc tgatggagat gggagctcat gggacccata agcttcatct 2280

ggtttcgctg gtctgagtga tttggttgcc ctcacaattt ggtggtggcg cctctgcagg 2340

acaaaataaa aataaacatt attattggtt ttatctgtta aaaaaacttc attttcccct 2400

cttaatgctt ggtctgcaac tagcccttcc ttcagagctc aagattcatt tgctctttcc 2460

cacagccaca gttcctgggt acagtccaag taggaattct ttctaaaagg ggtctgcatg 2520

gtcattttaa taaaatcaag tgtacaagga aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2580

aaaa 2584

<210> 118

<211> 698

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> transferrin

<400> 118

Met Arg Leu Ala Val Gly Ala Leu Leu Val Cys Ala Val Leu Gly Leu

1 5 10 15

Cys Leu Ala Val Pro Asp Lys Thr Val Arg Trp Cys Ala Val Ser Glu

20 25 30

His Glu Ala Thr Lys Cys Gln Ser Phe Arg Asp His Met Lys Ser Val

35 40 45

Ile Pro Ser Asp Gly Pro Ser Val Ala Cys Val Lys Lys Ala Ser Tyr

50 55 60

Leu Asp Cys Ile Arg Ala Ile Ala Ala Asn Glu Ala Asp Ala Val Thr

65 70 75 80

Leu Asp Ala Gly Leu Val Tyr Asp Ala Tyr Leu Ala Pro Asn Asn Leu

85 90 95

Lys Pro Val Val Ala Glu Phe Tyr Gly Ser Lys Glu Asp Pro Gln Thr

100 105 110

Phe Tyr Tyr Ala Val Ala Val Val Lys Lys Asp Ser Gly Phe Gln Met

115 120 125

Asn Gln Leu Arg Gly Lys Lys Ser Cys His Thr Gly Leu Gly Arg Ser

130 135 140

Ala Gly Trp Asn Ile Pro Ile Gly Leu Leu Tyr Cys Asp Leu Pro Glu

145 150 155 160

Pro Arg Lys Pro Leu Glu Lys Ala Val Ala Asn Phe Phe Ser Gly Ser

165 170 175

Cys Ala Pro Cys Ala Asp Gly Thr Asp Phe Pro Gln Leu Cys Gln Leu

180 185 190

Cys Pro Gly Cys Gly Cys Ser Thr Leu Asn Gln Tyr Phe Gly Tyr Ser

195 200 205

Gly Ala Phe Lys Cys Leu Lys Asp Gly Ala Gly Asp Val Ala Phe Val

210 215 220

Lys His Ser Thr Ile Phe Glu Asn Leu Ala Asn Lys Ala Asp Arg Asp

225 230 235 240

Gln Tyr Glu Leu Leu Cys Leu Asp Asn Thr Arg Lys Pro Val Asp Glu

245 250 255

Tyr Lys Asp Cys His Leu Ala Gln Val Pro Ser His Thr Val Val Ala

260 265 270

Arg Ser Met Gly Gly Lys Glu Asp Leu Ile Trp Glu Leu Leu Asn Gln

275 280 285

Ala Gln Glu His Phe Gly Lys Asp Lys Ser Lys Glu Phe Gln Leu Phe

290 295 300

Ser Ser Pro His Gly Lys Asp Leu Leu Phe Lys Asp Ser Ala His Gly

305 310 315 320

Phe Leu Lys Val Pro Pro Arg Met Asp Ala Lys Met Tyr Leu Gly Tyr

325 330 335

Glu Tyr Val Thr Ala Ile Arg Asn Leu Arg Glu Gly Thr Cys Pro Glu

340 345 350

Ala Pro Thr Asp Glu Cys Lys Pro Val Lys Trp Cys Ala Leu Ser His

355 360 365

His Glu Arg Leu Lys Cys Asp Glu Trp Ser Val Asn Ser Val Gly Lys

370 375 380

Ile Glu Cys Val Ser Ala Glu Thr Thr Glu Asp Cys Ile Ala Lys Ile

385 390 395 400

Met Asn Gly Glu Ala Asp Ala Met Ser Leu Asp Gly Gly Phe Val Tyr

405 410 415

Ile Ala Gly Lys Cys Gly Leu Val Pro Val Leu Ala Glu Asn Tyr Asn

420 425 430

Lys Ser Asp Asn Cys Glu Asp Thr Pro Glu Ala Gly Tyr Phe Ala Val

435 440 445

Ala Val Val Lys Lys Ser Ala Ser Asp Leu Thr Trp Asp Asn Leu Lys

450 455 460

Gly Lys Lys Ser Cys His Thr Ala Val Gly Arg Thr Ala Gly Trp Asn

465 470 475 480

Ile Pro Met Gly Leu Leu Tyr Asn Lys Ile Asn His Cys Arg Phe Asp

485 490 495

Glu Phe Phe Ser Glu Gly Cys Ala Pro Gly Ser Lys Lys Asp Ser Ser

500 505 510

Leu Cys Lys Leu Cys Met Gly Ser Gly Leu Asn Leu Cys Glu Pro Asn

515 520 525

Asn Lys Glu Gly Tyr Tyr Gly Tyr Thr Gly Ala Phe Arg Cys Leu Val

530 535 540

Glu Lys Gly Asp Val Ala Phe Val Lys His Gln Thr Val Pro Gln Asn

545 550 555 560

Thr Gly Gly Lys Asn Pro Asp Pro Trp Ala Lys Asn Leu Asn Glu Lys

565 570 575

Asp Tyr Glu Leu Leu Cys Leu Asp Gly Thr Arg Lys Pro Val Glu Glu

580 585 590

Tyr Ala Asn Cys His Leu Ala Arg Ala Pro Asn His Ala Val Val Thr

595 600 605

Arg Lys Asp Lys Glu Ala Cys Val His Lys Ile Leu Arg Gln Gln Gln

610 615 620

His Leu Phe Gly Ser Asn Val Thr Asp Cys Ser Gly Asn Phe Cys Leu

625 630 635 640

Phe Arg Ser Glu Thr Lys Asp Leu Leu Phe Arg Asp Asp Thr Val Cys

645 650 655

Leu Ala Lys Leu His Asp Arg Asn Thr Tyr Glu Lys Tyr Leu Gly Glu

660 665 670

Glu Tyr Val Lys Ala Val Gly Asn Leu Arg Lys Cys Ser Thr Ser Ser

675 680 685

Leu Leu Glu Ala Cys Thr Phe Arg Arg Pro

690 695

<210> 119

<211> 2808

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> transferrin cDNA

<400> 119

tggcaccgag cgagccgcga tgacaatggc tgcattgtgc ttcatgtccc ttcccatcaa 60

catttctgtg ctggactcct tccactcgcg ggtcgtctcc agagctcaga aaatgaggtg 120

atcagtggga cgagtaagga aggggggttg ggagaggggc gattgggcaa cccggctgca 180

caaacacggg aggtcaaaga ttgcgcccag cccgcccagg ccgggaatgg aataaaggga 240

cgcggggcgc cggaggctgc acagaagcga gtccgactgt gctcgctgct cagcgccgca 300

cccggaagat gaggctcgcc gtgggagccc tgctggtctg cgccgtcctg gggctgtgtc 360

tggctgtccc tgataaaact gtgagatggt gtgcagtgtc ggagcatgag gccactaagt 420

gccagagttt ccgcgaccat atgaaaagcg tcattccatc cgatggtccc agtgttgctt 480

gtgtgaagaa agcctcctac cttgattgca tcagggccat tgcggcaaac gaagcggatg 540

ctgtgacact ggatgcaggt ttggtgtatg atgcttacct ggctcccaat aacctgaagc 600

ctgtggtggc agagttctat gggtcaaaag aggatccaca gactttctat tatgctgttg 660

ctgtggtgaa gaaggatagt ggcttccaga tgaaccagct tcgaggcaag aagtcctgcc 720

acacgggtct aggcaggtcc gctgggtgga acatccccat aggcttactt tactgtgact 780

tacctgagcc acgtaaacct cttgagaaag cagtggccaa tttcttctcg ggcagctgtg 840

ccccttgtgc ggatgggacg gacttccccc agctgtgtca actgtgtcca gggtgtggct 900

gctccaccct taaccaatac ttcggctact cgggagcctt caagtgtctg aaggatggtg 960

ctggggatgt ggcctttgtc aagcactcga ctatatttga gaacttggca aacaaggctg 1020

acagggacca gtatgagctg ctttgcctgg acaacacccg gaagccggta gatgaataca 1080

aggactgcca cttggcccag gtcccttctc ataccgtcgt ggcccgaagt atgggcggca 1140

aggaggactt gatctgggag cttctcaacc aggcccagga acattttggc aaagacaaat 1200

caaaagaatt ccaactattc agctctcctc atgggaagga cctgctgttt aaggactctg 1260

cccacgggtt tttaaaagtc ccccccagga tggatgccaa gatgtacctg ggctatgagt 1320

atgtcactgc catccggaat ctacgggaag gcacatgccc agaagcccca acagatgaat 1380

gcaagcctgt gaagtggtgt gcgctgagcc accacgagag gctcaagtgt gatgagtgga 1440

gtgttaacag tgtagggaaa atagagtgtg tatcagcaga gaccaccgaa gactgcatcg 1500

ccaagatcat gaatggagaa gctgatgcca tgagcttgga tggagggttt gtctacatag 1560

cgggcaagtg tggtctggtg cctgtcttgg cagaaaacta caataagagc gataattgtg 1620

aggatacacc agaggcaggg tattttgctg tagcagtggt gaagaaatca gcttctgacc 1680

tcacctggga caatctgaaa ggcaagaagt cctgccatac ggcagttggc agaaccgctg 1740

gctggaacat ccccatgggc ctgctctaca ataagatcaa ccactgcaga tttgatgaat 1800

ttttcagtga aggttgtgcc cctgggtcta agaaagactc cagtctctgt aagctgtgta 1860

tgggctcagg cctaaacctg tgtgaaccca acaacaaaga gggatactac ggctacacag 1920

gcgctttcag gtgtctggtt gagaagggag atgtggcctt tgtgaaacac cagactgtcc 1980

cacagaacac tgggggaaaa aaccctgatc catgggctaa gaatctgaat gaaaaagact 2040

atgagttgct gtgccttgat ggtaccagga aacctgtgga ggagtatgcg aactgccacc 2100

tggccagagc cccgaatcac gctgtggtca cacggaaaga taaggaagct tgcgtccaca 2160

agatattacg tcaacagcag cacctatttg gaagcaacgt aactgactgc tcgggcaact 2220

tttgtttgtt ccggtcggaa accaaggacc ttctgttcag agatgacaca gtatgtttgg 2280

ccaaacttca tgacagaaac acatatgaaa aatacttagg agaagaatat gtcaaggctg 2340

ttggtaacct gagaaaatgc tccacctcat cactcctgga agcctgcact ttccgtagac 2400

cttaaaatct cagaggtagg gctgccacca aggtgaagat gggaacgcag atgatccatg 2460

agtttgccct ggtttcactg gcccaagtgg tttgtgctaa ccacgtctgt cttcacagct 2520

ctgtgttgcc atgtgtgctg aacaaaaaat aaaaattatt attgatttta tatttcaaaa 2580

actccattct ttcctaaata ttttcaacaa aggatttctt tatgcattct gcctaaatac 2640

ctatgcaact gagcccttcc ttctcagctc aagattcgtc tggtctttcc ctacagcttt 2700

gtgtgtgcca tggccacatc tcctgggtac agttcaagga gacatctttt ctaaaagggt 2760

ctgcgtgatc attaaaatat aatcaaatgt aaaaaaaaaa aaaaaaaa 2808

<210> 120

<211> 607

<212> PRT

<213> cattle

<220>

<221> variants

<222> (0)...(0)

<223> serum albumin

<400> 120

Met Lys Trp Val Thr Phe Ile Ser Leu Leu Leu Leu Phe Ser Ser Ala

1 5 10 15

Tyr Ser Arg Gly Val Phe Arg Arg Asp Thr His Lys Ser Glu Ile Ala

20 25 30

His Arg Phe Lys Asp Leu Gly Glu Glu His Phe Lys Gly Leu Val Leu

35 40 45

Ile Ala Phe Ser Gln Tyr Leu Gln Gln Cys Pro Phe Asp Glu His Val

50 55 60

Lys Leu Val Asn Glu Leu Thr Glu Phe Ala Lys Thr Cys Val Ala Asp

65 70 75 80

Glu Ser His Ala Gly Cys Glu Lys Ser Leu His Thr Leu Phe Gly Asp

85 90 95

Glu Leu Cys Lys Val Ala Ser Leu Arg Glu Thr Tyr Gly Asp Met Ala

100 105 110

Asp Cys Cys Glu Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Ser

115 120 125

His Lys Asp Asp Ser Pro Asp Leu Pro Lys Leu Lys Pro Asp Pro Asn

130 135 140

Thr Leu Cys Asp Glu Phe Lys Ala Asp Glu Lys Lys Phe Trp Gly Lys

145 150 155 160

Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro Glu

165 170 175

Leu Leu Tyr Tyr Ala Asn Lys Tyr Asn Gly Val Phe Gln Glu Cys Cys

180 185 190

Gln Ala Glu Asp Lys Gly Ala Cys Leu Leu Pro Lys Ile Glu Thr Met

195 200 205

Arg Glu Lys Val Leu Thr Ser Ser Ala Arg Gln Arg Leu Arg Cys Ala

210 215 220

Ser Ile Gln Lys Phe Gly Glu Arg Ala Leu Lys Ala Trp Ser Val Ala

225 230 235 240

Arg Leu Ser Gln Lys Phe Pro Lys Ala Glu Phe Val Glu Val Thr Lys

245 250 255

Leu Val Thr Asp Leu Thr Lys Val His Lys Glu Cys Cys His Gly Asp

260 265 270

Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile Cys

275 280 285

Asp Asn Gln Asp Thr Ile Ser Ser Lys Leu Lys Glu Cys Cys Asp Lys

290 295 300

Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu Lys Asp Ala

305 310 315 320

Ile Pro Glu Asn Leu Pro Pro Leu Thr Ala Asp Phe Ala Glu Asp Lys

325 330 335

Asp Val Cys Lys Asn Tyr Gln Glu Ala Lys Asp Ala Phe Leu Gly Ser

340 345 350

Phe Leu Tyr Glu Tyr Ser Arg Arg His Pro Glu Tyr Ala Val Ser Val

355 360 365

Leu Leu Arg Leu Ala Lys Glu Tyr Glu Ala Thr Leu Glu Glu Cys Cys

370 375 380

Ala Lys Asp Asp Pro His Ala Cys Tyr Ser Thr Val Phe Asp Lys Leu

385 390 395 400

Lys His Leu Val Asp Glu Pro Gln Asn Leu Ile Lys Gln Asn Cys Asp

405 410 415

Gln Phe Glu Lys Leu Gly Glu Tyr Gly Phe Gln Asn Ala Leu Ile Val

420 425 430

Arg Tyr Thr Arg Lys Val Pro Gln Val Ser Thr Pro Thr Leu Val Glu

435 440 445

Val Ser Arg Ser Leu Gly Lys Val Gly Thr Arg Cys Cys Thr Lys Pro

450 455 460

Glu Ser Glu Arg Met Pro Cys Thr Glu Asp Tyr Leu Ser Leu Ile Leu

465 470 475 480

Asn Arg Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Glu Lys Val

485 490 495

Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe Ser

500 505 510

Ala Leu Thr Pro Asp Glu Thr Tyr Val Pro Lys Ala Phe Asp Glu Lys

515 520 525

Leu Phe Thr Phe His Ala Asp Ile Cys Thr Leu Pro Asp Thr Glu Lys

530 535 540

Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Leu Lys His Lys Pro

545 550 555 560

Lys Ala Thr Glu Glu Gln Leu Lys Thr Val Met Glu Asn Phe Val Ala

565 570 575

Phe Val Asp Lys Cys Cys Ala Ala Asp Asp Lys Glu Ala Cys Phe Ala

580 585 590

Val Glu Gly Pro Lys Leu Val Val Ser Thr Gln Thr Ala Leu Ala

595 600 605

<210> 121

<211> 1883

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> serum albumin cDNA

<400> 121

gtcgactttg gcacaatgaa gtgggtgact tttatttctc ttctccttct cttcagctct 60

gcttattcca ggggtgtgtt tcgtcgagat acacacaaga gtgagattgc tcatcggttt 120

aaagatttgg gagaagaaca ttttaaaggc ctggtactga ttgccttttc tcagtatctc 180

cagcagtgtc catttgatga gcatgtaaaa ttagtgaacg aactaactga gtttgcaaaa 240

acatgtgttg ctgatgagtc ccatgccggc tgtgaaaagt cacttcacac tctctttgga 300

gatgaattgt gtaaagttgc atcccttcgt gaaacctatg gtgacatggc tgactgctgt 360

gagaaacaag agcctgaaag aaatgaatgc ttcctgagcc acaaagatga tagcccagac 420

ctccctaaat tgaaaccaga ccccaatact ttgtgtgatg agtttaaggc agatgaaaag 480

aagttttggg gaaaatacct atacgaaatt gctagaagac atccctactt ttatgcacca 540

gaactccttt actatgctaa taaatataat ggagtttttc aagaatgctg ccaagctgaa 600

gataaaggtg cctgcctgct accaaagatt gaaactatga gagaaaaagt actgacttca 660

tctgccagac agagactcag gtgtgccagt attcaaaaat ttggagaaag agctttaaaa 720

gcatggtcag tagctcgcct gagccagaaa tttcccaagg ctgagtttgt agaagttacc 780

aagctagtga cagatctcac aaaagtccac aaggaatgct gccatggtga cctacttgaa 840

tgcgcagatg acagggcaga tcttgccaag tacatatgtg ataatcaaga tacaatctcc 900

agtaaactga aggaatgctg tgataagcct ttgttggaaa aatcccactg cattgctgag 960

gtagaaaaag atgccatacc tgaaaacctg cccccattaa ctgctgactt tgctgaagat 1020

aaggatgttt gcaaaaacta tcaggaagca aaagatgcct tcctgggctc gtttttgtat 1080

gaatattcaa gaaggcatcc tgaatatgct gtctcagtgc tattgagact tgccaaggaa 1140

tatgaagcca cactggagga atgctgtgcc aaagatgatc cacatgcatg ctattccaca 1200

gtgtttgaca aacttaagca tcttgtggat gagcctcaga atttaatcaa acaaaactgt 1260

gaccaattcg aaaaacttgg agagtatgga ttccaaaatg cgctcatagt tcgttacacc 1320

aggaaagtac cccaagtgtc aactccaact ctcgtggagg tttcaagaag cctaggaaaa 1380

gtgggtacta ggtgttgtac aaagccggaa tcagaaagaa tgccctgtac tgaagactat 1440

ctgagcttga tcctgaaccg gttgtgcgtg ctgcatgaga agacaccagt gagtgaaaaa 1500

gtcaccaagt gctgcacaga gtcattggtg aacagacggc catgtttctc tgctctgaca 1560

cctgatgaaa catatgtacc caaagccttt gatgagaaat tgttcacctt ccatgcagat 1620

atatgcacac ttcccgatac tgagaaacaa atcaagaaac aaactgcact tgttgagctg 1680

ttgaaacaca agcccaaggc aacagaggaa caactgaaaa ccgtcatgga gaattttgtg 1740

gcttttgtag acaagtgctg tgcagctgat gacaaagagg cctgctttgc tgtggagggt 1800

ccaaaacttg ttgtttcaac tcaaacagcc ttagcctaaa cacgacacaa ccacaggcat 1860

ctcagcctac cctgagagtc gac 1883

<210> 122

<211> 607

<212> PRT

<213> goat

<220>

<221> variants

<222> (0)...(0)

<223> serum albumin

<400> 122

Met Lys Trp Val Thr Phe Ile Ser Leu Leu Leu Leu Phe Ser Ser Ala

1 5 10 15

Tyr Ser Arg Gly Val Phe Arg Arg Asp Thr His Lys Ser Glu Ile Ala

20 25 30

His Arg Phe Asn Asp Leu Gly Glu Glu Asn Phe Gln Gly Leu Val Leu

35 40 45

Ile Ala Phe Ser Gln Tyr Leu Gln Gln Cys Pro Phe Asp Glu His Val

50 55 60

Lys Leu Val Lys Glu Leu Thr Glu Phe Ala Lys Thr Cys Val Ala Asp

65 70 75 80

Glu Ser His Ala Gly Cys Asp Lys Ser Leu His Thr Leu Phe Gly Asp

85 90 95

Glu Leu Cys Lys Val Ala Thr Leu Arg Glu Thr Tyr Gly Asp Met Ala

100 105 110

Asp Cys Cys Glu Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Lys

115 120 125

His Lys Asp Asp Ser Pro Asp Leu Pro Lys Leu Lys Pro Glu Pro Asp

130 135 140

Thr Leu Cys Ala Glu Phe Lys Ala Asp Glu Lys Lys Phe Trp Gly Lys

145 150 155 160

Tyr Leu Tyr Glu Val Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro Glu

165 170 175

Leu Leu Tyr Tyr Ala Asn Lys Tyr Asn Gly Val Phe Gln Glu Cys Cys

180 185 190

Gln Ala Glu Asp Lys Gly Ala Cys Leu Leu Pro Lys Ile Glu Thr Met

195 200 205

Arg Glu Lys Val Leu Ala Ser Ser Ala Arg Gln Arg Leu Arg Cys Ala

210 215 220

Ser Ile Gln Lys Phe Gly Glu Arg Ala Leu Lys Ala Trp Ser Val Ala

225 230 235 240

Arg Leu Ser Gln Lys Phe Pro Lys Ala Asp Phe Thr Asp Val Thr Lys

245 250 255

Ile Val Thr Asp Leu Thr Lys Val His Lys Glu Cys Cys His Gly Asp

260 265 270

Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile Cys

275 280 285

Asp His Gln Asp Thr Leu Ser Ser Lys Leu Lys Glu Cys Cys Asp Lys

290 295 300

Pro Val Leu Glu Lys Ser His Cys Ile Ala Glu Ile Asp Lys Asp Ala

305 310 315 320

Val Pro Glu Asn Leu Pro Pro Leu Thr Ala Asp Phe Ala Glu Asp Lys

325 330 335

Glu Val Cys Lys Asn Tyr Gln Glu Ala Lys Asp Val Phe Leu Gly Ser

340 345 350

Phe Leu Tyr Glu Tyr Ser Arg Arg His Pro Glu Tyr Ala Val Ser Val

355 360 365

Leu Leu Arg Leu Ala Lys Glu Tyr Glu Ala Thr Leu Glu Asp Cys Cys

370 375 380

Ala Lys Glu Asp Pro His Ala Cys Tyr Ala Thr Val Phe Asp Lys Leu

385 390 395 400

Lys His Leu Val Asp Glu Pro Gln Asn Leu Ile Lys Lys Asn Cys Glu

405 410 415

Leu Phe Glu Lys His Gly Glu Tyr Gly Phe Gln Asn Ala Leu Ile Val

420 425 430

Arg Tyr Thr Arg Lys Ala Pro Gln Val Ser Thr Pro Thr Leu Val Glu

435 440 445

Ile Ser Arg Ser Leu Gly Lys Val Gly Thr Lys Cys Cys Ala Lys Pro

450 455 460

Glu Ser Glu Arg Met Pro Cys Thr Glu Asp Tyr Leu Ser Leu Ile Leu

465 470 475 480

Asn Arg Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Glu Lys Val

485 490 495

Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe Ser

500 505 510

Asp Leu Thr Leu Asp Glu Thr Tyr Val Pro Lys Pro Phe Asp Gly Glu

515 520 525

Ser Phe Thr Phe His Ala Asp Ile Cys Thr Leu Pro Asp Thr Glu Lys

530 535 540

Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Leu Lys His Lys Pro

545 550 555 560

Lys Ala Thr Asp Glu Gln Leu Lys Thr Val Met Glu Asn Phe Val Ala

565 570 575

Phe Val Asp Lys Cys Cys Ala Ala Asp Asp Lys Glu Gly Cys Phe Leu

580 585 590

Leu Glu Gly Pro Lys Leu Val Ala Ser Thr Gln Ala Ala Leu Ala

595 600 605

<210> 123

<211> 2072

<212> DNA

<213> goat

<220>

<221> misc_feature

<222> (0)...(0)

<223> serum albumin cDNA

<400> 123

tttccctcta ttcatactat cttttctatc aaccccacaa acctttggca caatgaagtg 60

ggtgactttt atttcccttc tccttctctt cagctctgct tattccaggg gtgtgtttcg 120

tcgagataca cacaaaagtg agattgctca tcggtttaat gatttgggag aagaaaattt 180

tcaaggcctg gtgctgattg ccttttctca gtatctccag cagtgtccat ttgatgaaca 240

tgtaaaatta gtgaaggaac taactgagtt tgcaaaaaca tgtgttgctg atgagtcaca 300

tgccggttgt gataagtcac ttcacactct ctttggagat gaattgtgta aagttgcaac 360

ccttcgcgaa acctatggtg acatggccga ctgctgtgag aaacaagagc ctgaaagaaa 420

tgaatgcttc ctgaaacaca aagatgatag cccagacctc cctaaactga aaccagagcc 480

cgatactttg tgtgccgagt ttaaggcaga tgaaaagaag ttttggggaa aatacctata 540

cgaagttgcc agaagacatc cctactttta tgcaccagaa ctcctttact atgccaataa 600

atataatgga gtttttcaag aatgctgcca agctgaagat aaaggtgcct gcctactacc 660

aaagattgaa actatgagag aaaaagtact ggcttcatct gccagacaga gactcaggtg 720

tgccagtatt caaaaattcg gagaaagagc tttaaaagca tggtcagtag ctcgcctgag 780

ccagaaattt cccaaggctg actttacaga tgttaccaag atagtgacag atctcactaa 840

ggtccacaag gagtgctgcc atggcgacct acttgaatgc gcagacgaca gggcagatct 900

tgccaagtac atatgtgatc atcaagacac actctccagt aaactgaagg aatgctgtga 960

taagcctgtg ttggaaaaat cccactgcat tgctgagata gataaagatg ccgtgcctga 1020

aaacctgccc ccattaactg ctgactttgc tgaagataag gaggtttgca aaaactatca 1080

ggaagcaaaa gacgtcttcc tgggctcgtt tttgtatgaa tattcaagaa ggcatcctga 1140

gtatgctgtc tcagtgctgt tgagacttgc caaggaatat gaagccacac tggaggactg 1200

ctgtgctaaa gaagatccac atgcatgcta tgccacagtg tttgacaaac ttaagcatct 1260

tgtggatgag cctcagaatt taatcaaaaa aaactgtgag ctatttgaaa aacatggaga 1320

gtatggattc caaaatgcgc tcatagttcg ttacaccagg aaagcacccc aagtgtcaac 1380

tccaactctg gtggagattt caagaagcct aggaaaagtg ggcactaagt gttgtgcaaa 1440

gcctgaatca gaaagaatgc cctgtaccga agactatctg agcttgatcc tgaaccggtt 1500

gtgcgtgttg cacgagaaga caccagtgag tgaaaaagtc accaagtgct gcacagagtc 1560

attggtgaac agacggccat gtttctctga tctgacactt gacgaaacat atgtacccaa 1620

acccttcgat ggtgaatctt tcaccttcca tgcagatata tgcacacttc ctgatactga 1680

gaaacaaatc aagaaacaaa ctgcacttgt tgagctgttg aaacacaagc ccaaggcaac 1740

agatgaacaa ctgaaaaccg ttatggagaa ttttgtggct tttgtagaca agtgctgcgc 1800

agctgatgac aaagaaggct gctttcttct ggagggtcca aaacttgttg cttcaactca 1860

agcagcctta gcctaaacac gacacaacca caagcatctc agcctaccct gagagtaaga 1920

cggaagaaga gaaatgaaaa ctcagagctt attcatctgt tcttcttttc tgttggtgtt 1980

aaacctacac cctctctaaa gaacataaat ttctttaaat attttgcttc ttttgtttgt 2040

gctacaatta ataaaaaatg aaaagactct aa 2072

<210> 124

<211> 609

<212> PRT

<213> Intelligent people

<220>

<221> variants

<222> (0)...(0)

<223> serum albumin

<400> 124

Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala

1 5 10 15

Tyr Ser Arg Gly Val Phe Arg Arg Asp Ala His Lys Ser Glu Val Ala

20 25 30

His Arg Phe Lys Asp Leu Gly Glu Glu Asn Phe Lys Ala Leu Val Leu

35 40 45

Ile Ala Phe Ala Gln Tyr Leu Gln Gln Cys Pro Phe Glu Asp His Val

50 55 60

Lys Leu Val Asn Glu Val Thr Glu Phe Ala Lys Thr Cys Val Ala Asp

65 70 75 80

Glu Ser Ala Glu Asn Cys Asp Lys Ser Leu His Thr Leu Phe Gly Asp

85 90 95

Lys Leu Cys Thr Val Ala Thr Leu Arg Glu Thr Tyr Gly Glu Met Ala

100 105 110

Asp Cys Cys Ala Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Gln

115 120 125

His Lys Asp Asp Asn Pro Asn Leu Pro Arg Leu Val Arg Pro Glu Val

130 135 140

Asp Val Met Cys Thr Ala Phe His Asp Asn Glu Glu Thr Phe Leu Lys

145 150 155 160

Lys Tyr Leu Tyr Glu Ile Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro

165 170 175

Glu Leu Leu Phe Phe Ala Lys Arg Tyr Lys Ala Ala Phe Thr Glu Cys

180 185 190

Cys Gln Ala Ala Asp Lys Ala Ala Cys Leu Leu Pro Lys Leu Asp Glu

195 200 205

Leu Arg Asp Glu Gly Lys Ala Ser Ser Ala Lys Gln Arg Leu Lys Cys

210 215 220

Ala Ser Leu Gln Lys Phe Gly Glu Arg Ala Phe Lys Ala Trp Ala Val

225 230 235 240

Ala Arg Leu Ser Gln Arg Phe Pro Lys Ala Glu Phe Ala Glu Val Ser

245 250 255

Lys Leu Val Thr Asp Leu Thr Lys Val His Thr Glu Cys Cys His Gly

260 265 270

Asp Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile

275 280 285

Cys Glu Asn Gln Asp Ser Ile Ser Ser Lys Leu Lys Glu Cys Cys Glu

290 295 300

Lys Pro Leu Leu Glu Lys Ser His Cys Ile Ala Glu Val Glu Asn Asp

305 310 315 320

Glu Met Pro Ala Asp Leu Pro Ser Leu Ala Ala Asp Phe Val Glu Ser

325 330 335

Lys Asp Val Cys Lys Asn Tyr Ala Glu Ala Lys Asp Val Phe Leu Gly

340 345 350

Met Phe Leu Tyr Glu Tyr Ala Arg Arg His Pro Asp Tyr Ser Val Val

355 360 365

Leu Leu Leu Arg Leu Ala Lys Thr Tyr Lys Thr Thr Leu Glu Lys Cys

370 375 380

Cys Ala Ala Ala Asp Pro His Glu Cys Tyr Ala Lys Val Phe Asp Glu

385 390 395 400

Phe Lys Pro Leu Val Glu Glu Pro Gln Asn Leu Ile Lys Gln Asn Cys

405 410 415

Glu Leu Phe Glu Gln Leu Gly Glu Tyr Lys Phe Gln Asn Ala Leu Leu

420 425 430

Val Arg Tyr Thr Lys Lys Val Pro Gln Val Ser Thr Pro Thr Leu Val

435 440 445

Glu Val Ser Arg Asn Leu Gly Lys Val Gly Ser Lys Cys Cys Lys His

450 455 460

Pro Glu Ala Lys Arg Met Pro Cys Ala Glu Asp Tyr Leu Ser Val Val

465 470 475 480

Leu Asn Gln Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Asp Arg

485 490 495

Val Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe

500 505 510

Ser Ala Leu Glu Val Asp Glu Thr Tyr Val Pro Lys Glu Phe Asn Ala

515 520 525

Glu Thr Phe Thr Phe His Ala Asp Ile Cys Thr Leu Ser Glu Lys Glu

530 535 540

Arg Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Val Lys His Lys

545 550 555 560

Pro Lys Ala Thr Lys Glu Gln Leu Lys Ala Val Met Asp Asp Phe Ala

565 570 575

Ala Phe Val Glu Lys Cys Cys Lys Ala Asp Asp Lys Glu Thr Cys Phe

580 585 590

Ala Glu Glu Gly Lys Lys Leu Val Ala Ala Ser Arg Ala Ala Leu Gly

595 600 605

Leu

<210> 125

<211> 2176

<212> DNA

<213> Intelligent people

<220>

<221> misc_feature

<222> (0)...(0)

<223> serum albumin cDNA

<400> 125

tctcttctgt caaccccacg cgcctttggc acaatgaagt gggtaacctt tatttccctt 60

ctttttctct ttagctcggc ttattccagg ggtgtgtttc gtcgagatgc acacaagagt 120

gaggttgctc atcggtttaa agatttggga gaagaaaatt tcaaagcctt agtgttgatt 180

gcctttgctc agtatcttca gcagtgtcca tttgaagatc atgtaaaatt agtgaatgaa 240

gtaactgaat ttgcaaaaac atgtgttgct gatgagtcag ctgaaaattg tgacaaatca 300

cttcataccc tttttggaga caaattatgc acagttgcaa ctcttcgtga aacctatggt 360

gaaatggctg actgctgtgc aaaacaagaa cctgagagaa atgaatgctt cttgcaacac 420

aaagatgaca acccaaacct cccccgattg gtgagaccag aggttgatgt gatgtgcact 480

gcttttcatg acaatgaaga gacatttttg aaaaaatact tatatgaaat tgccagaaga 540

catccttact tctatgcccc ggaactcctt ttctttgcta aaaggtataa agctgctttt 600

acagaatgtt gccaagctgc tgataaggct gcctgcctgt tgccaaagct cgatgaactt 660

cgggatgaag ggaaggcttc gtctgccaaa cagaggctca agtgtgccag tctccaaaaa 720

tttggagaaa gagctttcaa agcatgggca gtagctcgcc tgagccagag atttcccaaa 780

gctgagtttg cagaagtttc caagttagtg acagatctta ccaaagtcca cacggaatgc 840

tgccatggag atctgcttga atgtgctgat gacagggcgg accttgccaa gtatatctgt 900

gaaaatcaag attcgatctc cagtaaactg aaggaatgct gtgaaaaacc tctgttggaa 960

aaatcccact gcattgccga agtggaaaat gatgagatgc ctgctgactt gccttcatta 1020

gctgctgatt ttgttgaaag taaggatgtt tgcaaaaact atgctgaggc aaaggatgtc 1080

ttcctgggca tgtttttgta tgaatatgca agaaggcatc ctgattactc tgtcgtgctg 1140

ctgctgagac ttgccaagac atataaaacc actctagaga agtgctgtgc cgctgcagat 1200

cctcatgaat gctatgccaa agtgttcgat gaatttaaac ctcttgtgga agagcctcag 1260

aatttaatca aacaaaattg tgagcttttt gagcagcttg gagagtacaa attccagaat 1320

gcgctattag ttcgttacac caagaaagta ccccaagtgt caactccaac tcttgtagag 1380

gtctcaagaa acctaggaaa agtgggcagc aaatgttgta aacatcctga agcaaaaaga 1440

atgccctgtg cagaagacta tctatccgtg gtcctgaacc agttatgtgt gttgcatgag 1500

aaaacgccag taagtgacag agtcaccaaa tgctgcacag aatccttggt gaacaggcga 1560

ccatgctttt cagctctgga agtcgatgaa acatacgttc ccaaagagtt taatgctgaa 1620

acattcacct tccatgcaga tatatgcaca ctttctgaga aggagagaca aatcaagaaa 1680

caaactgcac ttgttgagct tgtgaaacac aagcccaagg caacaaaaga gcaactgaaa 1740

gctgttatgg atgatttcgc agcttttgta gagaagtgct gcaaggctga cgataaggag 1800

acctgctttg ccgaggaggg taaaaaactt gttgctgcaa gtcgagctgc cttaggctta 1860

taacatcaca tttaaaagca tctcagccta ccatgagaat aagagaaaga aaatgaagat 1920

caaaagctta ttcatctgtt tttctttttc gttggtgtaa agccaacacc ctgtctaaaa 1980

aacataaatt tctttaatca ttttgcctct tttctctgtg cttcaattaa taaaaaatgg 2040

aaagaatcta atagagtggt acagcactgt tatttttcaa agatgtgttg ctatcctgaa 2100

aattctgtag gttctgtgga agttccagtg ttctctctta ttccacttcg gtagaggatt 2160

tctagtttct gtgggc 2176

<210> 126

<211> 607

<212> PRT

<213> domestic sheep

<220>

<221> variants

<222> (0)...(0)

<223> serum albumin

<400> 126

Met Lys Trp Val Thr Phe Ile Ser Leu Leu Leu Leu Phe Ser Ser Ala

1 5 10 15

Tyr Ser Arg Gly Val Phe Arg Arg Asp Thr His Lys Ser Glu Ile Ala

20 25 30

His Arg Phe Asn Asp Leu Gly Glu Glu Asn Phe Gln Gly Leu Val Leu

35 40 45

Ile Ala Phe Ser Gln Tyr Leu Gln Gln Cys Pro Phe Asp Glu His Val

50 55 60

Lys Leu Val Lys Glu Leu Thr Glu Phe Ala Lys Thr Cys Val Ala Asp

65 70 75 80

Glu Ser His Ala Gly Cys Asp Lys Ser Leu His Thr Leu Phe Gly Asp

85 90 95

Glu Leu Cys Lys Val Ala Thr Leu Arg Glu Thr Tyr Gly Asp Met Ala

100 105 110

Asp Cys Cys Glu Lys Gln Glu Pro Glu Arg Asn Glu Cys Phe Leu Asn

115 120 125

His Lys Asp Asp Ser Pro Asp Leu Pro Lys Leu Lys Pro Glu Pro Asp

130 135 140

Thr Leu Cys Ala Glu Phe Lys Ala Asp Glu Lys Lys Phe Trp Gly Lys

145 150 155 160

Tyr Leu Tyr Glu Val Ala Arg Arg His Pro Tyr Phe Tyr Ala Pro Glu

165 170 175

Leu Leu Tyr Tyr Ala Asn Lys Tyr Asn Gly Val Phe Gln Glu Cys Cys

180 185 190

Gln Ala Glu Asp Lys Gly Ala Cys Leu Leu Pro Lys Ile Asp Ala Met

195 200 205

Arg Glu Lys Val Leu Ala Ser Ser Ala Arg Gln Arg Leu Arg Cys Ala

210 215 220

Ser Ile Gln Lys Phe Gly Glu Arg Ala Leu Lys Ala Trp Ser Val Ala

225 230 235 240

Arg Leu Ser Gln Lys Phe Pro Lys Ala Asp Phe Thr Asp Val Thr Lys

245 250 255

Ile Val Thr Asp Leu Thr Lys Val His Lys Glu Cys Cys His Gly Asp

260 265 270

Leu Leu Glu Cys Ala Asp Asp Arg Ala Asp Leu Ala Lys Tyr Ile Cys

275 280 285

Asp His Gln Asp Ala Leu Ser Ser Lys Leu Lys Glu Cys Cys Asp Lys

290 295 300

Pro Val Leu Glu Lys Ser His Cys Ile Ala Glu Val Asp Lys Asp Ala

305 310 315 320

Val Pro Glu Asn Leu Pro Pro Leu Thr Ala Asp Phe Ala Glu Asp Lys

325 330 335

Glu Val Cys Lys Asn Tyr Gln Glu Ala Lys Asp Val Phe Leu Gly Ser

340 345 350

Phe Leu Tyr Glu Tyr Ser Arg Arg His Pro Glu Tyr Ala Val Ser Val

355 360 365

Leu Leu Arg Leu Ala Lys Glu Tyr Glu Ala Thr Leu Glu Asp Cys Cys

370 375 380

Ala Lys Glu Asp Pro His Ala Cys Tyr Ala Thr Val Phe Asp Lys Leu

385 390 395 400

Lys His Leu Val Asp Glu Pro Gln Asn Leu Ile Lys Lys Asn Cys Glu

405 410 415

Leu Phe Glu Lys His Gly Glu Tyr Gly Phe Gln Asn Ala Leu Ile Val

420 425 430

Arg Tyr Thr Arg Lys Ala Pro Gln Val Ser Thr Pro Thr Leu Val Glu

435 440 445

Ile Ser Arg Ser Leu Gly Lys Val Gly Thr Lys Cys Cys Ala Lys Pro

450 455 460

Glu Ser Glu Arg Met Pro Cys Thr Glu Asp Tyr Leu Ser Leu Ile Leu

465 470 475 480

Asn Arg Leu Cys Val Leu His Glu Lys Thr Pro Val Ser Glu Lys Val

485 490 495

Thr Lys Cys Cys Thr Glu Ser Leu Val Asn Arg Arg Pro Cys Phe Ser

500 505 510

Asp Leu Thr Leu Asp Glu Thr Tyr Val Pro Lys Pro Phe Asp Glu Lys

515 520 525

Phe Phe Thr Phe His Ala Asp Ile Cys Thr Leu Pro Asp Thr Glu Lys

530 535 540

Gln Ile Lys Lys Gln Thr Ala Leu Val Glu Leu Leu Lys His Lys Pro

545 550 555 560

Lys Ala Thr Asp Glu Gln Leu Lys Thr Val Met Glu Asn Phe Val Ala

565 570 575

Phe Val Asp Lys Cys Cys Ala Ala Asp Asp Lys Glu Gly Cys Phe Val

580 585 590

Leu Glu Gly Pro Lys Leu Val Ala Ser Thr Gln Ala Ala Leu Ala

595 600 605

<210> 127

<211> 2089

<212> DNA

<213> domestic sheep

<220>

<221> misc_feature

<222> (0)...(0)

<223> serum albumin cDNA

<400> 127

gaattccttt tttttctttt ctatcaaccc cacaaacctt tggcacaatg aagtgggtga 60

cttttatttc ccttctcctt ctcttcagct ctgcttattc caggggtgtg tttcgtcgag 120

atacacacaa gagtgagatt gctcatcggt ttaatgattt gggagaagaa aattttcaag 180

gcctggtgct gattgccttt tctcagtatc tccagcagtg tccatttgac gaacatgtaa 240

aattagtgaa ggagctaact gagtttgcaa aaacatgtgt tgctgatgag tcacatgccg 300

gttgtgataa gtcacttcac actctctttg gagatgaatt gtgtaaagtt gcaacccttc 360

gcgaaaccta tggtgacatg gccgactgct gtgagaaaca agagcctgaa agaaatgaat 420

gcttcctgaa tcacaaagat gatagcccag acctccctaa actgaaacca gagcccgata 480

ctttgtgtgc cgagtttaag gcagatgaaa agaagttttg gggaaaatac ctatacgaag 540

ttgccagaag acatccctac ttttatgcac cagaactcct ttactatgct aataaatata 600

atggagtttt tcaagaatgc tgccaagctg aagataaagg tgcctgccta ctaccaaaga 660

ttgacgctat gagagaaaaa gtactggctt catctgccag acagagactc aggtgtgcca 720

gtattcaaaa attcggagaa agagctttaa aagcatggtc agtagctcgc ctgagccaga 780

aatttcccaa ggctgacttt acagatgtta ccaagatagt gacagatctc actaaggtcc 840

acaaggagtg ttgccatggt gacctgcttg aatgcgcaga cgacagggca gatcttgcca 900

agtacatatg tgatcatcaa gacgcactct ccagtaaact gaaggaatgc tgtgataagc 960

ctgtgttgga aaaatcccac tgcattgctg aggtagataa agatgccgtg cctgaaaacc 1020

tgcccccatt aactgctgac tttgctgaag ataaggaggt ttgcaaaaac tatcaggaag 1080

caaaagacgt cttcctgggc tcgtttttgt atgaatattc aagaaggcat cctgagtatg 1140

ctgtctcagt gctattgaga cttgccaagg aatatgaagc cacactggag gactgctgtg 1200

ccaaagaaga tccacatgcc tgctatgcca cagtgtttga caaacttaag catcttgtgg 1260

atgagcctca gaatttaatc aaaaaaaact gtgagctatt cgaaaaacat ggagagtatg 1320

gattccaaaa tgcgctcata gttcgttaca ccaggaaagc accccaagtg tcaactccaa 1380

ctctggtgga gatttcaaga agcctaggaa aagtgggcac taagtgttgt gcaaagcctg 1440

aatcagaaag aatgccctgt accgaagact atctgagctt gatcctgaac cggttgtgcg 1500

tgttgcatga gaagacacca gtgagtgaaa aagtcaccaa gtgctgcacg gagtcattgg 1560

tgaacagacg gccatgtttc tctgatctga cacttgacga aacatatgta cccaaaccct 1620

tcgatgagaa atttttcacc ttccatgcag atatatgcac acttcctgat actgagaaac 1680

aaatcaagaa acaaactgca cttgttgagc tgttgaaaca caagcccaag gcaacagatg 1740

aacaactgaa aaccgttatg gagaattttg tggcttttgt agacaagtgc tgtgcagctg 1800

atgacaaaga aggctgcttt gttctggagg gtccaaaact tgttgcttca actcaagcag 1860

ccttagccta aacacgacac aaccacaagc atctcagcct accctgagag tgagacgaaa 1920

aaagagaaat gaaaactcag agcttattca tctgttcttc ttttcgggtg ttggtgttaa 1980

acctacaccc tctctaaaga acataaattt ctttaaatat tttgcttctt ttgtttgtgc 2040

tacaattaat aaaaaatgaa aagactctaa aaaaaaaaaa aaggaattc 2089

<210> 128

<211> 42

<212> PRT

<213> Artificial sequence

<220>

<223> joint

<400> 128

Gly Gly Cys Thr Cys Ala Gly Gly Ala Thr Cys Ala Gly Gly Gly Thr

1 5 10 15

Cys Gly Ala Ala Ala Ala Gly Ala Gly Gly Cys Thr Cys Ala Gly Gly

20 25 30

Ala Thr Cys Ala Gly Gly Gly Thr Cys Gly

35 40

<210> 129

<211> 943

<212> DNA

<213> Artificial sequence

<220>

<223> AOX1 promoter

<400> 129

aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca tccgacatcc acaggtccat 60

tctcacacat aagtgccaaa cgcaacagga ggggatacac tagcagcaga ccgttgcaaa 120

cgcaggacct ccactcctct tctcctcaac acccactttt gccatcgaaa aaccagccca 180

gttattgggc ttgattggag ctcgctcatt ccaattcctt ctattaggct actaacacca 240

tgactttatt agcctgtcta tcctggcccc cctggcgagg ttcatgtttg tttatttccg 300

aatgcaacaa gctccgcatt acacccgaac atcactccag atgagggctt tctgagtgtg 360

gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg acagtttaaa cgctgtcttg 420

gaacctaata tgacaaaagc gtgatctcat ccaagatgaa ctaagtttgg ttcgttgaaa 480

tgctaacggc cagttggtca aaaagaaact tccaaaagtc ggcataccgt ttgtcttgtt 540

tggtattgat tgacgaatgc tcaaaaataa tctcattaat gcttagcgca gtctctctat 600

cgcttctgaa ccccggtgca cctgtgccga aacgcaaatg gggaaacacc cgctttttgg 660

atgattatgc attgtctcca cattgtatgc ttccaagatt ctggtgggaa tactgctgat 720

agcctaacgt tcatgatcaa aatttaactg ttctaacccc tacttgacag caatatataa 780

acagaaggaa gctgccctgt cttaaacctt tttttttatc atcattatta gcttactttc 840

ataattgcga ctggttccaa ttgacaagct tttgatttta acgactttta acgacaactt 900

gagaagatca aaaaacaact aattattgaa agaattcaaa acg 943

<210> 130

<211> 486

<212> DNA

<213> Artificial sequence

<220>

<223> GAP1 promoter

<400> 130

tttttgtaga aatgtcttgg tgtcctcgtc caatcaggta gccatctctg aaatatctgg 60

ctccgttgca actccgaacg acctgctggc aacgtaaaat tctccggggt aaaacttaaa 120

tgtggagtaa tggaaccaga aacgtctctt cccttctctc tccttccacc gcccgttacc 180

gtccctagga aattttactc tgctggagag cttcttctac ggcccccttg cagcaatgct 240

cttcccagca ttacgttgcg ggtaaaacgg aggtcgtgta cccgacctag cagcccaggg 300

atggaaaagt cccggccgtc gctggcaata atagcgggcg gacgcatgtc atgagattat 360

tggaaaccac cagaatcgaa tataaaaggc gaacaccttt cccaattttg gtttctcctg 420

acccaaagac tttaaattta atttatttgt ccctatttca atcaattgaa caactatcaa 480

aacaca 486

<210> 131

<211> 441

<212> DNA

<213> Artificial sequence

<220>

<223> PGK1 promoter

<400> 131

agaacggaaa ggaatatatt tactgccgat cgcattttgg cctcaaataa atcttgagct 60

tttggacata gattatatgt tctttcttgg aagctctttc agctaatagt gaagtgtttc 120

ctactaagga tcgcctccaa acgttccaac tacgggcgga ggttgcaaag aaaacgggtc 180

tctcagcgaa ttgttctcat ccatgagtga gtcctctccg tcctttcctc gcgcctggca 240

ataaagcctc cttcggagga gctccgtcta gagaataatt gctgcctttc tgactttcgg 300

actagcgcca accgcgaacc acaccaccac accatcactg tcacccgtca tagttcatcc 360

ctctctcctt ataaagcatc taataggttc cacaattgtt tgccacaaaa atctcttagc 420

atagcccaat tgattacgaa a 441

<210> 132

<211> 57

<212> DNA

<213> Artificial sequence

<220>

<223> MF α T Signal peptide

<400> 132

atgaggtttc cttctatctt cacggcagtt cttttcgctg catcttccgc attggct 57

<210> 133

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> misc_feature

<222> (0)...(0)

<223> Signal peptide of alpha-S2

<220>

<223> kanamycin resistance gene

<400> 133

atgaagttct tcatcttcac ttgtttgttg gctgttgctt tggct 45

<210> 134

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<223> signal peptide of ost beta

<400> 134

atgagacagg tttggttctc ctggatcgtt ggtttgttct tgtgtttctt caacgtttcc 60

tccgctgct 69

<210> 135

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<223> beta Signal peptide

<400> 135

atgaaggttt tgatcttggc ttgtttggtt gctttggctt tggct 45

<210> 136

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<223> ost alpha-S2 signal peptide

<400> 136

atgagacagg tttggttcag ttggatagtc ggtttattcc tatgcttttt taatgtctca 60

tccgccgct 69

<210> 137

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of alpha-S1

<400> 137

atgaagttgt tgatcttgac ttgtttggtt gctgttgctt tggct 45

<210> 138

<211> 63

<212> DNA

<213> Artificial sequence

<220>

<223> k Signal peptide

<400> 138

atgatgaagt ccttcttctt ggttgttact atcttggctt tgactttgcc attcttgggt 60

gct 63

<210> 139

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of Ost alpha-S1

<400> 139

atgaggcaag tttggtttag ttggattgtt ggactgttcc tgtgcttttt caacgtgtca 60

tccgctgcc 69

<210> 140

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of Ost k

<400> 140

atgcgacagg tatggttttc ttggattgtg gggttgtttc tatgtttctt taacgtttct 60

tctgctgca 69

<210> 141

<211> 48

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of INU

<400> 141

atgaagttgg cttactcctt gttgttgcca ttggctggtg tttctgct 48

<210> 142

<211> 57

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of INV

<400> 142

atgttgttgc aggctttctt gttcttgttg gctggtttcg ctgctaagat ctctgct 57

<210> 143

<211> 489

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> beta-lactoglobulin

<400> 143

ttaatcgtca ctcagacaat gaaaggtttg gacatccaaa aagttgctgg aacttggtat 60

tcattggcaa tggctgcaag cgatatctcc ctgttagacg cccaatcagc tccactaaga 120

gtgtatgtgg aggagctgaa gcccactcca gaaggtgatc ttgaaattct gttgcaaaaa 180

tgggaaaatg gtgagtgtgc tcagaaaaag attatcgccg agaaaaccaa gattccggct 240

gtattcaaga tagatgctct gaacgagaat aaggtgttgg tcctcgatac agactacaaa 300

aagtaccttc tattctgtat ggaaaattcg gcagaacctg agcaatcctt ggcttgtcaa 360

tgcttggtta gaactccaga agttgacgac gaggctttgg aaaagtttga taaagccctg 420

aaagccttgc ctatgcatat tcgtttaagt tttaacccaa cacaattgga ggaacagtgt 480

cacatttaa 489

<210> 144

<211> 630

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein beta

<400> 144

agagagttgg aagagttgaa cgttccaggt gagatcgttg agtctttgtc ctcttcagaa 60

gagtccatca ctagaatcaa caagaagatc gagaagttcc agtccgagga acaacaacaa 120

actgaggacg agttgcagga caagattcac ccattcgctc aaactcagtc cttggtttac 180

ccattcccag gtccaattcc aaactccttg ccacagaaca tcccaccatt gactcagact 240

ccagttgttg ttccaccatt cttgcagcca gaggttatgg gtgtttccaa ggttaaggaa 300

gctatggctc caaagcacaa agagatgcca ttcccaaagt acccagttga gccattcact 360

gagtcccagt ccttgacttt gactgacgtt gagaacttgc acttgccatt gcctttgttg 420

caatcctgga tgcaccaacc acatcaacca ttgccaccaa ctgttatgtt cccaccacaa 480

tccgttttgt ccttgtccca atccaaggtt ttgccagttc cacagaaggc tgttccatac 540

cctcaaagag acatgccaat ccaggctttc ttgttgtacc aagagccagt tttgggtcca 600

gttagaggtc cattccctat catcgtttga 630

<210> 145

<211> 624

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2

<400> 145

aagaacacta tggaacacgt ttcatcctcc gaagagtcca tcatctccca agagacttac 60

aagcaagaga agaacatgga catcaaccca tccaaagaaa acttgtgttc cactttctgt 120

aaagaggttg ttagaaacgc taacgaggaa gagtactcca tcggttcctc atctgaagaa 180

tctgctgagg ttgctactga agaggttaag atcactgttg acgacaagca ctaccagaag 240

gctttgaacg agatcaacca gttctacaga aaattcccac aatacttgca gtacttgtac 300

cagggtccaa tcgttttgaa cccatgggac caggttaaga gaaacgctgt tcctatcact 360

ccaactttga acagagagca gttgtccact tccgaagaga actccaagaa aactgttgac 420

atggaatcca ctgaggtttt cactaagaaa actaagttga ctgaggaaga aaagaacaga 480

ttgaacttct tgaagaagat ctcccagaga taccagaagt tcgctttgcc acagtacttg 540

aaaacagttt accagcacca aaaggctatg aagccatgga tccagccaaa gactaaggtt 600

atcccatacg ttagatactt gtga 624

<210> 146

<211> 624

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S2K 113E variants

<400> 146

aagaacacta tggaacacgt ttcatcctcc gaagagtcca tcatctccca agagacttac 60

aagcaagaga agaacatgga catcaaccca tccaaagaaa acttgtgttc cactttctgt 120

aaagaggttg ttagaaacgc taacgaggaa gagtactcca tcggttcctc atctgaagaa 180

tctgctgagg ttgctactga agaggttaag atcactgttg acgacaagca ctaccagaag 240

gctttgaacg agatcaacca gttctacaga aaattcccac aatacttgca gtacttgtac 300

cagggtccaa tcgttttgaa cccatgggac caggttaagg aaaacgctgt tcctatcact 360

ccaactttga acagagagca gttgtccact tccgaagaga actccaagaa aactgttgac 420

atggaatcca ctgaggtttt cactaagaaa actaagttga ctgaggaaga aaagaacaga 480

ttgaacttct tgaagaagat ctcccagaga taccagaagt tcgctttgcc acagtacttg 540

aaaacagttt accagcacca aaaggctatg aagccatgga tccagccaaa gactaaggtt 600

atcccatacg ttagatactt gtga 624

<210> 147

<211> 600

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein alpha-S1

<400> 147

agacctaagc acccaatcaa gcaccaaggt ttgccacaag aggttttgaa cgagaacttg 60

ttgagattct tcgttgctcc attcccagag gttttcggta aagagaaggt taacgagttg 120

tccaaggaca tcggttctga atccactgag gaccaggcta tggaagatat caagcagatg 180

gaagctgagt ccatctcctc atccgaagag atcgttccaa actccgttga gcagaagcac 240

atccagaaag aggacgttcc atccgagaga tacttgggtt acttggagca gttgttgaga 300

ttgaagaagt acaaggttcc acagttggaa atcgttccta attccgctga agaaagattg 360

cactccatga aggaaggtat ccacgctcag cagaaagaac ctatgatcgg tgttaatcaa 420

gagttggctt acttctaccc agagttgttc agacagttct accagttgga cgcttaccca 480

tctggtgctt ggtactacgt tccattgggt actcagtaca ctgacgctcc atctttctcc 540

gacattccaa acccaattgg ttccgagaac tccggtaaga ctactatgcc attgtggtaa 600

<210> 148

<211> 510

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> Casein k

<400> 148

caagagcaaa atcaagagca gccaatcaga tgtgagaagg acgagagatt cttctcagac 60

aagatcgcta agtacatccc aatccagtac gttttgtcca gatacccatc ctacggtttg 120

aactactacc agcagaagcc agttgctttg atcaacaacc agttcttgcc atacccttac 180

tacgctaagc cagctgctgt tagatctcct gctcaaatct tgcagtggca ggttttgtct 240

aacactgttc cagctaagtc ctgtcaggct cagccaacta ctatggctag acatccacat 300

ccacacttgt ccttcatggc tatcccacca aagaagaacc aggacaagac tgagatccca 360

actatcaaca ctatcgcttc cggtgagcca acttccactc caactattga agctgttgag 420

tccactgttg ctactttgga agcttctcca gaggttattg aatccccacc agagatcaac 480

acagttcagg ttacttccac tgctgtttaa 510

<210> 149

<211> 279

<212> DNA

<213> Artificial sequence

<220>

<223> AOX1 terminator sequence

<400> 149

ggttaaaggg gcggccgctc aagaggatgt cagaatgcca tttgcctgag agatgcaggc 60

ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt ttgtcatttt 120

gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagcagatg aatatcttgt 180

ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc ccactcctct 240

tcagagtaca gaagattaag tgaaaccttc gtttgtgcg 279

<210> 150

<211> 271

<212> DNA

<213> Artificial sequence

<220>

<223> CYC1 terminator sequence

<400> 150

atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc 60

gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg 120

ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg 180

tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct 240

ttaatttgcg gcccctcacc tgcacgcaaa a 271

<210> 151

<211> 375

<212> DNA

<213> Artificial sequence

<220>

<223> bleomycin resistance gene

<400> 151

atggctaaac tcacctctgc tgttccagtc ctgactgctc gtgatgttgc tggtgctgtt 60

gagttctgga ctgatagact cggtttctcc cgtgacttcg tagaggacga ctttgccggt 120

gttgtacgtg acgacgttac cctgttcatc tccgcagttc aggaccaggt tgtgccagac 180

aacactctgg catgggtatg ggttcgtggt ctggacgaac tgtacgctga gtggtctgag 240

gtcgtgtcta ccaacttccg tgatgcatct ggtccagcta tgaccgagat cggtgaacag 300

ccctggggtc gtgagtttgc actgcgtgat ccagctggta actgcgtgca tttcgtcgca 360

gaagaacagg actaa 375

<210> 152

<211> 816

<212> DNA

<213> Artificial sequence

<220>

<223> kanamycin resistance gene

<400> 152

atgagccata ttcaacggga aacgtcttgc tcgaggccgc gattaaattc caacatggat 60

gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc 120

tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc 180

gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct 240

cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg 300

atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt 360

gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct 420

tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg 480

gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa 540

gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca 600

cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc 660

ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct 720

ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa 780

ttgcagtttc atttgatgct cgatgagttt ttctaa 816

<210> 153

<211> 939

<212> DNA

<213> Artificial sequence

<220>

<223> promoter AOX1

<400> 153

gatctaacat ccaaagacga aaggttgaat gaaacctttt tgccatccga catccacagg 60

tccattctca cacataagtg ccaaacgcaa caggagggga tacactagca gcagaccgtt 120

gcaaacgcag gacctccact cctcttctcc tcaacaccca cttttgccat cgaaaaacca 180

gcccagttat tgggcttgat tggagctcgc tcattccaat tccttctatt aggctactaa 240

caccatgact ttattagcct gtctatcctg gcccccctgg cgaggttcat gtttgtttat 300

ttccgaatgc aacaagctcc gcattacacc cgaacatcac tccagatgag ggctttctga 360

gtgtggggtc aaatagtttc atgttcccca aatggcccaa aactgacagt ttaaacgctg 420

tcttggaacc taatatgaca aaagcgtgat ctcatccaag atgaactaag tttggttcgt 480

tgaaatgcta acggccagtt ggtcaaaaag aaacttccaa aagtcggcat accgtttgtc 540

ttgtttggta ttgattgacg aatgctcaaa aataatctca ttaatgctta gcgcagtctc 600

tctatcgctt ctgaaccccg gtgcacctgt gccgaaacgc aaatggggaa acacccgctt 660

tttggatgat tatgcattgt ctccacattg tatgcttcca agattctggt gggaatactg 720

ctgatagcct aacgttcatg atcaaaattt aactgttcta acccctactt gacagcaata 780

tataaacaga aggaagctgc cctgtcttaa accttttttt ttatcatcat tattagctta 840

ctttcataat tgcgactggt tccaattgac aagcttttga ttttaacgac ttttaacgac 900

aacttgagaa gatcaaaaaa caactaatta ttcgaaacg 939

<210> 154

<211> 267

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of alpha mating factor

<400> 154

atgagatttc cttcaatttt tactgctgtt ttattcgcag catcctccgc attagctgct 60

ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc tgtcatcggt 120

tactcagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa cagcacaaat 180

aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga agaaggggta 240

tctctcgaga aaagagaggc tgaagct 267

<210> 155

<211> 66

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of OST1

<400> 155

atgagacaag tttggttctc atggattgtt ggtttattct tgtgtttctt caacgtttct 60

tccgct 66

<210> 156

<211> 57

<212> DNA

<213> Artificial sequence

<220>

<223> Signal peptide of alpha-lactalbumin

<400> 156

atgatgtcat ttgtttcttt gttgttggtt ggtatcctgt tccacgccac tcaagct 57

<210> 157

<211> 372

<212> DNA

<213> cattle

<220>

<221> misc_feature

<222> (0)...(0)

<223> alpha-lactalbumin

<400> 157

gagcaattga ccaagtgtga agttttcaga gagttgaagg atttgaaggg ttacggtggt 60

gtttctcttc cagaatgggt ttgtaccgct ttccacactt ccggatacga tacacaagct 120

atcgttcaaa acaacgactc caccgagtac ggtttgttcc aaatcaacaa caagatttgg 180

tgtaaagacg atcaaaaccc tcactcttcc aacatttgca acatctcttg tgataagttc 240

ttggatgatg atctcaccga tgatatcatg tgtgttaaga agattttgga caaagttgga 300

atcaactact ggttggctca caaggctctt tgttcagaga agttggacca atggctatgt 360

gagaagttgt aa 372

<210> 158

<211> 350

<212> DNA

<213> Artificial sequence

<220>

<223> AOX1 terminator sequence

<400> 158

tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt 60

ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc 120

ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa 180

tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta 240

agtgacacgt tcgtttgtgc aagcttcaac gatgccaaaa gggtataata agcgtcattt 300

gcagcattgt gaagaaaact atgtggcaag ccaagcctgc gaagaatgta 350

<210> 159

<211> 810

<212> DNA

<213> Artificial sequence

<220>

<223> Synthesis of oligonucleotide

<400> 159

atgggtaagg aaaagactca cgtttccaga ccaagattga actctaacat ggacgctgac 60

ttgtacggtt acaagtgggc tagagacaac gttggtcaat ctggtgctac tatttacaga 120

ttgtacggta agccagacgc tccagagttg ttcttgaagc acggtaaggg ttctgttgct 180

aacgacgtta ctgacgagat ggttagattg aactggttga ctgagttcat gccattgcca 240

actattaagc acttcattag aactccagac gacgcttggt tgttgactac tgctattcca 300

ggtaagactg ctttccaagt tttggaggag tacccagact ctggtgagaa cattgttgac 360

gctttggctg ttttcttgag aagattgcac tctattccag tttgtaactg tccattcaac 420

tctgacagag ttttcagatt ggctcaagct caatccagaa tgaacaacgg tttggttgac 480

gcttctgact tcgacgacga gagaaacggt tggccagttg agcaagtttg gaaggagatg 540

cacaagttgt tgccattctc tccagactct gttgttactc acggtgactt ctctttggac 600

aacttgattt tcgacgaggg taagttgatt ggttgtattg acgttggtag agttggtatt 660

gctgacagat accaagactt ggctattttg tggaactgtt tgggtgagtt ctctccatct 720

ttgcaaaaga gattgttcca aaagtacggt attgacaacc cagacatgaa caagttgcaa 780

ttccacttga tgttggacga gttcttctaa 810

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Composition comprising casein and method for producing the same

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