阅读说明：本技术 一种快速去除皱纹使皮肤年轻化的混合物料的制备方法 (Preparation method of mixed material for rapidly removing wrinkles and rejuvenating skin ) 是由 朱瑜 于 2020-07-16 设计创作，主要内容包括：本发明公开了一种快速去除皱纹使皮肤年轻化的混合物料的制备方法,所述方法包括以下步骤：将外基质蛋白的编码基因克隆插入质粒pET-32a(+)中；通过CaCl2转化法转化至感受态细胞大肠杆菌细胞BL21(DE3)中；获得包含该重组质粒的基因工程菌；发酵及菌体收集：选取所述步骤S3中获得的阳性基因工程菌克隆,置于培养基中培养至菌体达目标浓度,然后收集菌体；纯化与透析：将所述步骤S4中收集到的菌体去除杂质后,进行蛋白纯化操作,再进行蛋白透析操作,即获得所述混合物料。本发明有益效果在于,遗传背景清楚、繁殖快、成本低、表达量高、表达产物容易纯化、稳定性好、抗污染能力强等。(The invention discloses a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin, which comprises the following steps: cloning and inserting the coding gene of the outer matrix protein into a plasmid pET-32a (+); transforming into competent cell Escherichia coli cell BL21(DE3) by CaCl2 transformation method; obtaining genetically engineered bacteria containing the recombinant plasmid; fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium; purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material. The invention has the advantages of clear genetic background, quick propagation, low cost, high expression quantity, easy purification of expression products, good stability, strong pollution resistance and the like.)

1. A method for preparing a skin rejuvenating composition for rapid wrinkle removal, said method comprising the steps of:

s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);

s2 passing through CaCl₂Transformation into competent cells by transformationEnterobacter cells BL21(DE 3);

s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;

s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;

s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.

2. The method of claim 1, wherein the matrix proteins include collagen type I, collagen type IV, and fibronectin.

3. Use of the mixture prepared according to claim 1 in the field of skin care products.

Technical Field

The invention belongs to the field of recombinant protein expression, and particularly relates to a preparation method of a mixed material for rapidly removing wrinkles and enabling skin to be young.

Background

Collagen is widely distributed in the body of multicellular animals. Collagen has been thought to function only as a supporting tissue protein. In recent years, research on collagen has been increasing, and due to its excellent biological properties and weak antigenicity, it can be used in the medical and health fields of burns, wounds, beauty, orthopedics, tissue repair, wound hemostasis, etc. after being specially treated, and this research has been successful. Therefore, it is necessary to research and develop the synthesized recombinant protein using the related art of genetically engineered bacteria.

Disclosure of Invention

The invention mainly aims to overcome the defects of the prior art and provide a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for preparing a mixed material for rapidly removing wrinkles to rejuvenate skin, the method comprising the steps of:

s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);

s2 was transformed into competent cell E.coli cell BL21(DE3) by CaCl2 transformation;

s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;

s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;

s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.

Preferably, the matrix proteins include collagen type I, collagen type IV, fibronectin.

The invention also provides application of the prepared mixed material in the field of skin care products.

The invention has the advantages of clear genetic background, quick propagation, low cost, high expression quantity, easy purification of expression products, good stability, strong pollution resistance and the like.

Detailed Description

The present invention will be further described below, and it should be noted that the following examples are provided to illustrate the detailed embodiments and specific procedures based on the technical solution, but the scope of the present invention is not limited to the examples.

The invention relates to a preparation method of a mixed material for rapidly removing wrinkles and rejuvenating skin, which comprises the following steps:

s1, cloning and inserting the coding gene of the extracellular matrix protein into a plasmid pET-32a (+);

s2 was transformed into competent cell E.coli cell BL21(DE3) by CaCl2 transformation;

s3, obtaining a genetically engineered bacterium containing the recombinant plasmid;

s4 fermentation and thallus collection: selecting the positive genetic engineering bacterium clone obtained in the step S3, placing the positive genetic engineering bacterium clone in a culture medium for culturing until the bacterium reaches the target concentration, and then collecting the bacterium;

s5 purification and dialysis: and (4) removing impurities from the thallus collected in the step S4, performing protein purification operation, and performing protein dialysis operation to obtain the mixed material.

Preferably, the matrix proteins include collagen type I, collagen type IV, fibronectin.

The invention also provides application of the prepared mixed material in the field of skin care products.

Various corresponding changes and modifications can be made by those skilled in the art based on the above technical solutions and concepts, and all such changes and modifications should be included in the protection scope of the present invention.