具体实施方式

本发明公开了大豆叶绿体膜蛋白基因或其蛋白的应用，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。

本发明提供了一种GmTic110基因，该GmTic110基因具有如下任一种核苷酸序列：

I)、如SEQ ID No.1所示的核苷酸序列；

II)、如SEQ ID No.1所示的核苷酸序列中经取代、缺失或添加一个或几个核苷酸且具有相同或相似功能的氨基酸序列；

III)、含SEQ ID No.1所示的核苷酸序列或其片段的载体。

本发明中，提供了GmTic110蛋白的氨基酸序列为如下序列之一：

I)、如SEQ ID No.2所示的氨基酸序列；

II)、如SEQ ID No.2所示的氨基酸序列中经取代、缺失或添加一个或几个氨基酸且具有相同或相似功能的氨基酸序列；

本发明中，编码GmTic110蛋白的核苷酸序列如SEQ ID No.3所示。

本发明中，提供了一种大豆GmTic110基因的突变体，其特征在于，其核苷酸序列为SEQ ID No.3所示的核酸序列中的第341位的G突变为A。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在调控叶绿素中的应用。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在调控叶绿体发育中的应用。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在调控光合作用中的应用。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在制备转基因植株中的应用。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在调控基因表达模式中的应用。

本发明提供了上述大豆GmTic110基因或含有该基因的生物材料在蛋白质亚细胞定位中的应用。

本发明还提供了包含CRISPR/Cas9系统在GmTic110基因上设计打靶位点，进行基因敲除载体构建，其骨架载体为VK005-04-soU6-2-GmUbi3。

含有上述表达载体的重组宿主为农杆菌，本发明所述的基因敲除的方法采用农杆菌介导法将含GmTic110基因靶位点的基因敲除载体转化入植物外植体。作为受体材料的外植体为胚轴细胞。

作为优选，基因敲除载体特异性引物序列如SEQ ID No.4和SEQ ID No.5所示。

SEQ ID No.4所示上游引物：GATTGCGGCGGCTGGATACGGCCT

SEQ ID No.5所示下游引物：AAACAGGCCGTATCCAGCCGCCGC

本发明中，所述植物为豆科植物；本发明中用以验证GmTic110功能的植物为豆科植物大豆，具体为大豆Williams 82。

所述编码GmTIC110蛋白的核酸分子包括编码GmTIC110蛋白的基因组DNA、cDNA、重组DNA或mRNA、hnRNA；或者与上述DNA、cDNA、重组DNA或mRNA呈反向互补的核酸分子。

上述核酸分子可根据实际需要进行修饰或优化，从而使基因表达更高效；例如，①可根据受体植物所偏爱的密码子，在保持本发明所述GmHY2基因的氨基酸序列的同时改变其密码子以符合受体植物的偏爱性。②或修饰邻近起始甲硫氨酸的基因序列，以使翻译有效起始；例如，利用在植物中已知的有效的序列进行修饰。③与各种植物表达的启动子连接，以利于其在植物中的表达；所述启动子可包括组成型、诱导型、时序调节、发育调节、化学调节、组织优选和组织特异性启动子；启动子的选择将随着表达时间和空间需要而变化，而且也取决于靶物种；④引入增强子序列，如内含子序列(例如来源于Adhl和bronzel)和病毒前导序列(例如来源于TMV、MCMV和AMV)。

本发明中，所述载体可为质粒、黏粒、噬菌体或病毒载体。所述的宿主可为真菌、细菌、藻类或细胞。

对于不含有GmTIC110的植物，可采用化学法、鸟枪法、微注射，电穿孔等方法，将GmTIC110的基因片段导入植物细胞，也可通过同源重组、锌指核酸酶、TALEN、CRISPR等方法将GmTIC110的基因片段导入植物细胞。

本发明的优益效果在于：

(1)GmTic110基因分离自大豆，作为大豆的内源基因，对大豆的基因工程改造和研究大豆分子调控机理具有深远意义。

(2)本发明研究表明，大豆编码叶绿体膜基因突变，可以改变大豆叶绿体的正常运输，可用于定向改造叶绿体功能，为大豆叶绿体发育研究提供基因资源，为大豆育种提供种质资源。

本发明采用的试材皆为普通市售品，皆可于市场购得。

下面结合实施例，进一步阐述本发明：

实施例1.大豆中GmTic110基因的分离和结构分析

(1)GmTic110基因的分离

从大豆品种Williams 82中提取总RNA，以总RNA为模板，Oligo(T)17为引物合成cDNA第一条链。以该cDNA第一条链为模板，分别利用正向引物

(5’-ATGAACCCTTCCACACTCACC-3’，如SEQ ID No.6所示)和反向引物(5’-CTAGAATACAAACTTCTCTTCCT-3’，如SEQ ID No.7所示)进行PCR扩增。获得一个长为2988bp的GmTic110基因cDNA片段，将该片段与pEasy-Blunt载体(全式金公司)相连，命名为Blunt-GmTic110。

获得的GmTic110基因，其基因序列如SEQ ID No.1所示，GmTic110基因CDS全长共2988bp；由该基因编码的蛋白质的氨基酸序列如SEQ ID No.2所示，共995个氨基酸。

(2)GmTic110基因结构分析

提取大豆品种Williams 82幼嫩叶片中的DNA，以该基因组DNA为模板，扩增获得GmTic110基因组片段，GmTic110基因组序列如SEQ ID No.3所示，全长共7280bp，包含15个外显子和14个内含子(如图1)。

实施例2.大豆GmTic110基因突变体的分离、筛选及表型鉴定

从本实验室EMS诱变的Williams82突变体库中，筛选到一个叶片黄化突变体命名为Gmtic110-1(GmTic110基因的核酸序列中的第341位的G突变为A，如SEQ ID No.8所示)。与野生型Williams82相比，Gmtic110-1突变体从幼苗期到成熟期呈现叶片黄化表型(图2)。同时根据基因型我们在突变体库中筛选到的等位突变体命名为Gmtic110-2，其中GmTic110-2突变体观察到叶片黄化表型(图2)。

(1)GmTic110基因调控叶绿素合成

由于突变体Gmtic110整个生育期表现出叶片黄化表型，通过分光光度计检测Williams82、Gmtic110-1和Gmtic110-2叶片的叶绿素和类胡萝卜素含量，结果表明突变体Gmtic110-1和Gmtic110-2中叶绿素a与叶绿素b的含量均显著低于野生型(图3)。这表明GmTic110基因突变影响了叶绿素a与叶绿素b的合成途径。

表1图3数据

(2)GmTic110基因调控大豆叶绿体发育

为了分析突变体叶片黄化是否会导致叶绿体结构变化，通过透射电镜观察了野生型Williams82和突变体Gmtic110-1、GmTic110-2叶绿体的超微结构。突变体Gmtic110叶绿体较野生型Williams82叶绿体的淀粉粒数目增加，基粒类囊体堆积变薄(图4)。由于类囊体膜上有光合色素组分，并且光合作用主要发生在类囊体上，突变体中叶绿素a和叶绿素b均明显减少，导致了基粒类囊体堆积变薄，从而导致光合速率的降低。

(3)GmTic110基因调控大豆光合作用

因Gmtic110突变体叶绿素含量显著降低，为了明确是否会对叶片的光合作用造成影响，分别检测了Williams82、Gmtic110-1和Gmtic110-2光合作用强度指标；结果表明突变体GmTic110-1和Gmtic110-2与Williams82野生型相比，其净光合速率降低，气孔导度降低、蒸腾速率降低，胞间CO2浓度较野生型升高(图5)。结果表明了突变体Gmtic110的叶绿素含量减少明显的影响了其叶片的光合作用能力。

表2图5原始数据

表3图5原始数据

表4图5原始数据

表5图5原始数据

实施例3.大豆GmTic110基因敲除载体的构建

在GmTic110基因跨膜结构域上设计了靶位点PCR扩增靶标片段，将打靶位点的上下游引物进行二聚化，并将其连接到VK005-04-soU6-2-GmUbi3敲除表达载体上。将敲除载体通过冻融法转化大肠菌株Trans5α，在大肠杆菌中的抗性为卡那霉素抗性，在植物体中的抗性为草铵膦(Bar)抗性。

实施例4.农杆菌介导法转化豆科植物

在本实施例中，通过农杆菌介导法转化大豆子叶节的方法，得到含有GmTic110靶位点的敲除表达载体的Williams 82的外植体。

(A)大豆外植体的获得

选取表面光滑、无破损、无病斑、无裂痕的Williams 82成熟大豆种子，以氯气方法灭菌14h。将灭菌后的种子在超净台上通风，使氯气完全挥发，在萌发培养基萌发处理6h。去掉大豆1/2胚轴，将大豆沿胚轴纵向切开，将剩余下胚轴作为农杆菌介导转化的受体材料。

(B)大豆转化

农杆菌介导法采用二次农杆菌侵染，在共培养基上于22℃暗培养5d；SI-I培养基中，于强光下培养7d；切去外植体大芽，在SI-II培养基中，于强光下培养14d；切去外植体子叶和胚轴，于SE培养基中，每14d继代一次；将约3cm丛生芽剪下，放入生根培养基中生根；将RM生根培养基中，根部生长发达的植株转入土中栽植。通过Bar抗性检测，筛选得到了16个抗性植株。在人工气候室中培养5个月后果荚开始成熟，6个月后收种完毕。

(C)可以遗传的转基因植株

将收获的T1代种子在人工气候室中种植，可以观察到叶片黄化的表型，与Gmtic110突变体表型一致(图6)。

实施例5.大豆GmTIC110蛋白的亚细胞定位

为了验证GmTic110蛋白亚细胞定位情况，将构建好的PUC19-GFP-GmTic110瞬时表达载体利用聚乙二醇介导法转化拟南芥叶片原生质体，利用尼康激光共聚焦显微镜进行共定位观察，GFP荧光定位在叶绿体内膜上(图7)。

比较例

对照组：Gmtic110突变体在外显子处缺失一个碱基导致第113个氨基酸开始发生突变，在第120个氨基酸发生提前终止，该突变发生在第二个跨膜结构域，导致叶片出现黄化表型；

实验组：本发明中Gmtic110-1突变体中基因第1个外显子发生一个碱基替换导致氨基酸替换，该突变位点发生在第二个跨膜结构域，Gmtic110-2突变体中基因第14个外显子发生一个碱基替换导致氨基酸替换，该位点发生在互作因子结合结构域。

本发明突变体材料叶片黄化表型比对照组的突变体材料显著，从叶绿素含量和光合作用指标上比较明显低于对照组。通过CRISPR/Cas9技术获得的Gmtic110-CR突变体的打靶位点在第一个跨膜结构域，分别获得了碱基缺失和替换情况的突变体。转基因敲除突变体材料的表型也要比对照组表型显著。

以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

序列表

<110> 中国科学院东北地理与农业生态研究所

<120> 大豆GmTic110基因的突变体及其应用

<130> MP21010838

<160> 9

<170> SIPOSequenceListing 1.0

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<212> DNA

<213> 人工序列(Artificial Sequence)

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acgacgttat ctacaatcta caaagaggct ctatcatctt cttcctcttt agttttttcg 60

ttaaacccta taataacaac cactccactc ttctctactc tccgcaacct tacgacgtcg 120

tctcaaccat gaacccttcc acactcaccc cttcccacac ccaccgtcct ctcctcctac 180

cctctccctt ccacaccaga cggcgtcgtt ttaaagtctc cctccctcgc tgttcctcct 240

cctccgccgc ctcctccccc cctcctccgc cacctccgcc gccgcagcga ccgcccaagg 300

acctcaaggg aatcgatgtc ctcgtcgaca agctctcgcc gccggccagg ctcgccacct 360

ccgccgtcat cgtcgccggc gccgcggcgg ctggatacgg cctcggctcc cgcttcggcg 420

gaagccgcta cgctgcgctc ggtggagctg tcgccctcgg cgcggccggc ggtgctgcgg 480

cgtacgctct gaacgcggct gctccgcaag tcgccgccgt gaatttgcac aactacgtcg 540

ccgcgttcga cgacccttcg aagctgaaga aggaagagat tgaggccatt gcgtccaagt 600

tagtttttag tttcaatttc taataataat aactgttcat ttttttttct ttgttaattt 660

gagtgtttgg ttttgaaatg gttaattttt caggtatggt gtgagcaagc aagatgaggc 720

attcaaaact gagatttgtc atatatattc cgagtgagtg acaaattatt attattgttg 780

ttgttgtact ggtaacgagt tattatttgt tacttgcatt tgtgtagtga taatgtgtta 840

tgatgatttg gctgtttaat tttataattg aaagattgat ttgtggcata acgtgttgtt 900

caggtttgta tcttctgtgc ttcctcctgg tggtgaggaa cttaaaggtg atgaggttga 960

taggattgtc agcttcaaaa attctttggg aattgatgac ccggatgctg ctgctatgca 1020

catggaggta ttttttgtgt ttgttgtgta tatatataca cctgattacc agagtagatc 1080

gagtgtgtat gtttattatg aggagtgaaa gcagttccgt gtcttactgg tttggtgctt 1140

gttgactccc tagattggga ggaaattttt caggcaaagg ttggaagttg gggatcgtga 1200

tgctgatgtt gagcaacgtc gggtaagttt gttgtgttgt gatgttatct taactgattg 1260

gttgcaaatt tgcaatagca tcatgggttt tagatgaata atggatactt ggagcttgac 1320

cattgttaga gtctccagtg caggcatttc aaaagttgat atatgtgtca aatcttgtat 1380

ttggagatgc atcgtctttc cttctacctt ggaagcgtgt attcaaggtc accgattccc 1440

aggtatttat ttcttattgg ttaactggta tgcctagctt gtgcaacgtg atattaatat 1500

gcaattagga tgagagattt ggagcatagg tttatgacta tttataggca tttgctcaaa 1560

aagggaagac ctaatttgct gcacaaaatg cattttgcca tgtttgtgat cagtgacatg 1620

ctggcagcat ttcttaataa ttcttatatg ttctcattgt catttcttat cttagtgttt 1680

ttgttatttt tcatttgact agttaactat aatctattta ccagattgaa gtagctgtac 1740

gtgacaatgc gcagcgattg tttgcttcca agctgaaatc agttggcaga ggtaattgtg 1800

tagtcagtaa ttcctcttgg tccttgtttg ctcttactgt aacaaagttg atcaactaac 1860

tcacatttac tcctcgagaa gttgtcataa cttcatacta ctatattact ggagaaatga 1920

ttggaaatga taatcttgtt ttgtaattgt gcatgcattt cttattaaat aaagccttta 1980

aattttgctt gtactcatct agtatgctgg attttgttct gtaattctct agaattacta 2040

tcatccaaac taacaaattc tttctatagc aatttttaga agttttaaat caggtttatt 2100

tcctaagtgt taaccaatga tttcttgcca gatattgatg cagaaaaact tgttgcacta 2160

agaaaagaac aacaattatg tcgcctttct gatgaggttc attttcatta tttcttcttt 2220

cataactttc aacttctctg ggttaaatcc ctgtctttgt cttgccatca aatagttcct 2280

tggtatagta taatcttttt attaattaaa ggaaattttc attgaagctt gctgagaact 2340

tgtttagaga tcacacaagg aaattggttg aggaaaatat ttcagaggca aatagaatac 2400

ttaaatctcg caccaaagca gtgtaagatt catggttttt ctaagtattc ttttttttaa 2460

gtattaaatt ttgaattctg ggcatgtttt tcttgccatt tgtttcatta gtgttactat 2520

tcatttctta atttttacaa aaatagaatt tgccatatct gatcatctgt aagtttcagt 2580

cctggagcta cgcaggcaat tgcagagctg gataaggtat tggcatttaa taatttactc 2640

atctcattta agaatcatcc agatgtggat cgctttgctc gtggtgttgg tccaatttct 2700

ttagtgggta agtgtctgtg actaggtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgta 2760

taacatgttc ttttctcttt ttaatcttaa ttcttgaagt ttaccacaat tcatttttgg 2820

cataggtgga gagtatgatg gtgacagaaa aatagaggac ttgaaactcc tttacagggc 2880

atatgtttca gatgctttgt ctggtgggcg catggaagat gataaggtaa ataaccaaga 2940

caattggata tgaactgtga acttcttaag atgtgttttc tcaatattct aatatatttt 3000

gaagttgagt ttgggttagt gttttccaga tgtatgtagt gattgaatag tatattttct 3060

aaaaaaattg gagtattcag cagctattta tatcatattg gagtattcac ttcccatggg 3120

tataatagag attgaaacct agcagtttag ccttagttat tctttattgt ttcttatgtt 3180

ttatctttct ttttgatgta gtaccaattg ccaaatcatg attgaacagg ctgttcagtg 3240

catgctacca tatccatatt gctcttattc ttatttttaa tatttcagtt agggcagtat 3300

attattttca gtacaagggt atttgtttac tgtatcaagc ctaatgaatt ttcacttatt 3360

gaagcatttt ttagtgttat aaaacaatgt cctgtctcct tgcttaacac taacaactta 3420

aagcttttag gtcaggtgat ttgttgactg ttaagatgta ctagcatacg attccccttt 3480

gaatatagtt gcatatgttc tctaatcttc ctttaatgag tcaacattca gataagcaaa 3540

gacacaacag tgtctcgaaa accatgatat ccattactaa agagttctta ttgatataag 3600

atatgaaact tgatctgtca aaacacttat tttttaatat aaaataaaaa atatatttta 3660

aatatattta ttattcgttg cagcttgctg cactaaatca gttacgaaat atattcggac 3720

tgggtaaacg tgaagccgag gccatttcac ttgatgttac gtcgaaggta tatcgcaaac 3780

gacttgcaca ggctgctgca gatggtgagt tagaaatggc tgatagcaaa gcagccttcc 3840

ttcaaaatct atgtgatgaa cttcattttg atccacaaaa ggccagtgaa cttcatgaag 3900

gtaaaactcg tcatgtttac agttagttgg aagttgacta tatttgaggc ctttattatt 3960

gaacacttta tgtgctgtcg gtttttattt atgtgcaatt cttaaatctt aaggttgtaa 4020

aatgtaaact tgtaaaaccc tttctacatg atgtgacttt tttttatata tatatttagt 4080

tataacacaa tctaatgata gtttattatt tttaagaaat ttatcggcaa aagcttcaga 4140

gatgtgtagc tgatggggag ctcaatgagg aggatgttgc tgctttgttg aggatgcgtg 4200

taatgctctg cataccgcaa cagattgttg aagcagctca ttcagatatc tgtggcagtt 4260

tgtttgaaaa ggtagtcaac gttacttttt cctatctaat tgtttggtcc ttcataagtg 4320

tcaaattagt gaaattacat atttcttgct atgcagaagt gcggtccatc catgcactgt 4380

aacaaactga taatgataat atgaatacat ggatgaacac taaaattgct ttacatatga 4440

atatgggttc aagatgatta tggaactatg gttatcaaac tcattctact ttctatttga 4500

atattggtag catgagaagt tacagctgtc acattttaat gtgttgataa acctgataat 4560

gctttcattt actggtagtg tggagatgga gaccatgaat ctactgcatt tttttttatc 4620

ttacaggaaa gatacactgt tgtactctgt actctgcatg ctaggttttg tctagaattt 4680

ctaaaatgtg tatctattgg aattgttact ttcaactgct tgtaatttat ttgggctttg 4740

tcttttcaat ttgtagacct gtatctactt gctataaaat ctgtttattg gtttgattat 4800

cttatatcaa agttgtacaa gtctagattc tatagtgctt agttgatggc agcgatatgt 4860

gaagaaatgg gtctgtttta ttttgatata tcatgagtct tgtttcaatc tccatcaaag 4920

atgttcaaaa ttttgttgag ttattactaa ttcatgtttc tccttttata ggttgtcaag 4980

gaggcaattg catcaggggt tgatggatat gatgctgaaa tccagaaatc agtaagaaaa 5040

gcagcacatg gcctgcgact tactagggag gttgctatgt ctattgcaag caaggcggtg 5100

agtatcaagt tatcagaaac tcaatggtga tttataaaga tttgaatagg tttaaatttc 5160

actgaggttt tttggaatta taggtaagga agatttttat taattacata aaacgtgcac 5220

gggcagctgg aaatcgtacc gagtctgcga aagaactgaa gaagatgata gccttcaaca 5280

ccttagttgt aactaatttg gtggaggaca ttaaagggga gtcaactgat atttcctctg 5340

aagaacctgt gaaagaggac attacacaaa ctgacgatga agaatgggaa tcacttcaga 5400

cactcaagaa aataagacca aataaagaac ttacggaaaa gttgggaaaa cctggtcaga 5460

cagaaattac tctgaaagat gatcttcctg aaagggatag gactgatctt tacaagacat 5520

acttacttta ttgtctaact ggtgaagtga caagggttcc atttggtgct cagatcacta 5580

ctaagaagga tgattctgag tatcttcttc taaatcagct tggtgggatc ctgggattga 5640

gtagtcaaga aatagtggaa gtgcacaggg gtctagctga gcaggctttt aggcaacagg 5700

ctgaggtaat tttagccgat ggacagttga caaaggccag ggtggagcag cttaataacc 5760

tccagaaaca agtaggctta cctcaagaat atgctcagaa aataatcaag agtataacca 5820

ctacaaaaat ggcagctgcc attgaaactg ctgtaactca agggaggctc aatatgaagc 5880

agataaggga acttaaggaa gctgatgttg atttagacag tatggtatct gagaacttga 5940

gagagaccct cttcaaaaaa actgttgatg acattttctc atccggtact ggagagtttg 6000

acactgagga agtatatgaa aaaatcccgt cagatctcaa cattaacaaa gagaaggcac 6060

gaggtgttgt tcatgagctt gcaaagggta gactatccaa ctctctgatt caggctgtct 6120

ctctactaag acagagaaat cagcagggag tggtaagtaa tattcaaagc ttcatgttca 6180

cttagtacta gtttggattt atgctggatt atctaaaatt acactcaaat actagatcac 6240

gtgaagtaaa tgttcataaa ttttttgttt cactttaaga aattgattct gagttgaagc 6300

aattttaggt aactttttct gttgaataaa aaaggtttat gttaagtttt attattaact 6360

tgctttcatt aaaaaaaaac ctccaaacat aaatcttttc acttcaaaat caattatgca 6420

aaatctactt cattcataat caattttgca aatgtttgtc ccaacacact tagtgcgtgt 6480

ttgtacaagc attggcaaat tgattttaaa ttgaattgat tttagttaaa ttgatttgaa 6540

agtgatgtga tttatgttta agtacaacat tttagatcca tcgtaaaaat tactcaaagg 6600

tcctttaact tagagtcaat tttggactta gaatcaattc tgaattcttc tttaacagga 6660

aaccaaacac gaaaatttat caaaaagcca ttctccaatg tgaaacaaaa cttgcactta 6720

ttcaaacagc ttttttacgt ttgtttgtca tggataatca tgtgtgattg cttcttttct 6780

tttgtgcagg tttcttcact caatgacttg ctggcatgtg acaaagcagt accctcacag 6840

ccagtttcat gggaagtgcc agaggagctt tctgatctat acaccatata cttgaagagt 6900

aatccaactc ctgagaattt gtctcgtttg caatatctgt tgggtataaa tgattccaca 6960

gctgctgctc ttagggagat tggagataga ttactcaata ctactgcgga ggaagagaag 7020

tttgtattct agtcttgtga taaaatgttt ggcattttag gcacactaga gaggctgcaa 7080

gtccccttaa ttttgccatt ttttattttc cattttgttt aacggtagca gaaaaagaat 7140

atcacgatgt ataaactttg gaatagagat gagtgcagtg ctgaaatctt agataatatt 7200

ttgtttgaga cagttgaatt aatcaattgc tattttttat gaatgaaaat aagttttcat 7260

aatcgtggac aagttttatc 7280

<210> 2

<211> 995

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 2

Met Asn Pro Ser Thr Leu Thr Pro Ser His Thr His Arg Pro Leu Leu

1 5 10 15

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

20 25 30

Pro Arg Cys Ser Ser Ser Ser Ala Ala Ser Ser Pro Pro Pro Pro Pro

35 40 45

Pro Pro Pro Pro Gln Arg Pro Pro Lys Asp Leu Lys Gly Ile Asp Val

50 55 60

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

65 70 75 80

Ile Val Ala Gly Ala Ala Ala Ala Gly Tyr Gly Leu Gly Ser Arg Phe

85 90 95

Gly Gly Ser Arg Tyr Ala Ala Leu Gly Gly Ala Val Ala Leu Gly Ala

100 105 110

Ala Gly Gly Ala Ala Ala Tyr Ala Leu Asn Ala Ala Ala Pro Gln Val

115 120 125

Ala Ala Val Asn Leu His Asn Tyr Val Ala Ala Phe Asp Asp Pro Ser

130 135 140

Lys Leu Lys Lys Glu Glu Ile Glu Ala Ile Ala Ser Lys Tyr Gly Val

145 150 155 160

Ser Lys Gln Asp Glu Ala Phe Lys Thr Glu Ile Cys His Ile Tyr Ser

165 170 175

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

180 185 190

Asp Glu Val Asp Arg Ile Val Ser Phe Lys Asn Ser Leu Gly Ile Asp

195 200 205

Asp Pro Asp Ala Ala Ala Met His Met Glu Ile Gly Arg Lys Phe Phe

210 215 220

Arg Gln Arg Leu Glu Val Gly Asp Arg Asp Ala Asp Val Glu Gln Arg

225 230 235 240

Arg Ala Phe Gln Lys Leu Ile Tyr Val Ser Asn Leu Val Phe Gly Asp

245 250 255

Ala Ser Ser Phe Leu Leu Pro Trp Lys Arg Val Phe Lys Val Thr Asp

260 265 270

Ser Gln Ile Glu Val Ala Val Arg Asp Asn Ala Gln Arg Leu Phe Ala

275 280 285

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

290 295 300

Ala Leu Arg Lys Glu Gln Gln Leu Cys Arg Leu Ser Asp Glu Leu Ala

305 310 315 320

Glu Asn Leu Phe Arg Asp His Thr Arg Lys Leu Val Glu Glu Asn Ile

325 330 335

Ser Glu Ala Asn Arg Ile Leu Lys Ser Arg Thr Lys Ala Val Pro Gly

340 345 350

Ala Thr Gln Ala Ile Ala Glu Leu Asp Lys Val Leu Ala Phe Asn Asn

355 360 365

Leu Leu Ile Ser Phe Lys Asn His Pro Asp Val Asp Arg Phe Ala Arg

370 375 380

Gly Val Gly Pro Ile Ser Leu Val Gly Gly Glu Tyr Asp Gly Asp Arg

385 390 395 400

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

405 410 415

Leu Ser Gly Gly Arg Met Glu Asp Asp Lys Leu Ala Ala Leu Asn Gln

420 425 430

Leu Arg Asn Ile Phe Gly Leu Gly Lys Arg Glu Ala Glu Ala Ile Ser

435 440 445

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

450 455 460

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

465 470 475 480

Asn Leu Cys Asp Glu Leu His Phe Asp Pro Gln Lys Ala Ser Glu Leu

485 490 495

His Glu Glu Ile Tyr Arg Gln Lys Leu Gln Arg Cys Val Ala Asp Gly

500 505 510

Glu Leu Asn Glu Glu Asp Val Ala Ala Leu Leu Arg Met Arg Val Met

515 520 525

Leu Cys Ile Pro Gln Gln Ile Val Glu Ala Ala His Ser Asp Ile Cys

530 535 540

Gly Ser Leu Phe Glu Lys Val Val Lys Glu Ala Ile Ala Ser Gly Val

545 550 555 560

Asp Gly Tyr Asp Ala Glu Ile Gln Lys Ser Val Arg Lys Ala Ala His

565 570 575

Gly Leu Arg Leu Thr Arg Glu Val Ala Met Ser Ile Ala Ser Lys Ala

580 585 590

Val Arg Lys Ile Phe Ile Asn Tyr Ile Lys Arg Ala Arg Ala Ala Gly

595 600 605

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

610 615 620

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

625 630 635 640

Asp Ile Ser Ser Glu Glu Pro Val Lys Glu Asp Ile Thr Gln Thr Asp

645 650 655

Asp Glu Glu Trp Glu Ser Leu Gln Thr Leu Lys Lys Ile Arg Pro Asn

660 665 670

Lys Glu Leu Thr Glu Lys Leu Gly Lys Pro Gly Gln Thr Glu Ile Thr

675 680 685

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

690 695 700

Tyr Leu Leu Tyr Cys Leu Thr Gly Glu Val Thr Arg Val Pro Phe Gly

705 710 715 720

Ala Gln Ile Thr Thr Lys Lys Asp Asp Ser Glu Tyr Leu Leu Leu Asn

725 730 735

Gln Leu Gly Gly Ile Leu Gly Leu Ser Ser Gln Glu Ile Val Glu Val

740 745 750

His Arg Gly Leu Ala Glu Gln Ala Phe Arg Gln Gln Ala Glu Val Ile

755 760 765

Leu Ala Asp Gly Gln Leu Thr Lys Ala Arg Val Glu Gln Leu Asn Asn

770 775 780

Leu Gln Lys Gln Val Gly Leu Pro Gln Glu Tyr Ala Gln Lys Ile Ile

785 790 795 800

Lys Ser Ile Thr Thr Thr Lys Met Ala Ala Ala Ile Glu Thr Ala Val

805 810 815

Thr Gln Gly Arg Leu Asn Met Lys Gln Ile Arg Glu Leu Lys Glu Ala

820 825 830

Asp Val Asp Leu Asp Ser Met Val Ser Glu Asn Leu Arg Glu Thr Leu

835 840 845

Phe Lys Lys Thr Val Asp Asp Ile Phe Ser Ser Gly Thr Gly Glu Phe

850 855 860

Asp Thr Glu Glu Val Tyr Glu Lys Ile Pro Ser Asp Leu Asn Ile Asn

865 870 875 880

Lys Glu Lys Ala Arg Gly Val Val His Glu Leu Ala Lys Gly Arg Leu

885 890 895

Ser Asn Ser Leu Ile Gln Ala Val Ser Leu Leu Arg Gln Arg Asn Gln

900 905 910

Gln Gly Val Val Ser Ser Leu Asn Asp Leu Leu Ala Cys Asp Lys Ala

915 920 925

Val Pro Ser Gln Pro Val Ser Trp Glu Val Pro Glu Glu Leu Ser Asp

930 935 940

Leu Tyr Thr Ile Tyr Leu Lys Ser Asn Pro Thr Pro Glu Asn Leu Ser

945 950 955 960

Arg Leu Gln Tyr Leu Leu Gly Ile Asn Asp Ser Thr Ala Ala Ala Leu

965 970 975

Arg Glu Ile Gly Asp Arg Leu Leu Asn Thr Thr Ala Glu Glu Glu Lys

980 985 990

Phe Val Phe

995

<210> 3

<211> 2988

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 3

atgaaccctt ccacactcac cccttcccac acccaccgtc ctctcctcct accctctccc 60

ttccacacca gacggcgtcg ttttaaagtc tccctccctc gctgttcctc ctcctccgcc 120

gcctcctccc cccctcctcc gccacctccg ccgccgcagc gaccgcccaa ggacctcaag 180

ggaatcgatg tcctcgtcga caagctctcg ccgccggcca ggctcgccac ctccgccgtc 240

atcgtcgccg gcgccgcggc ggctggatac ggcctcggct cccgcttcgg cggaagccgc 300

tacgctgcgc tcggtggagc tgtcgccctc ggcgcggccg gcggtgctgc ggcgtacgct 360

ctgaacgcgg ctgctccgca agtcgccgcc gtgaatttgc acaactacgt cgccgcgttc 420

gacgaccctt cgaagctgaa gaaggaagag attgaggcca ttgcgtccaa gtatggtgtg 480

agcaagcaag atgaggcatt caaaactgag atttgtcata tatattccga gtttgtatct 540

tctgtgcttc ctcctggtgg tgaggaactt aaaggtgatg aggttgatag gattgtcagc 600

ttcaaaaatt ctttgggaat tgatgacccg gatgctgctg ctatgcacat ggagattggg 660

aggaaatttt tcaggcaaag gttggaagtt ggggatcgtg atgctgatgt tgagcaacgt 720

cgggcatttc aaaagttgat atatgtgtca aatcttgtat ttggagatgc atcgtctttc 780

cttctacctt ggaagcgtgt attcaaggtc accgattccc agattgaagt agctgtacgt 840

gacaatgcgc agcgattgtt tgcttccaag ctgaaatcag ttggcagaga tattgatgca 900

gaaaaacttg ttgcactaag aaaagaacaa caattatgtc gcctttctga tgagcttgct 960

gagaacttgt ttagagatca cacaaggaaa ttggttgagg aaaatatttc agaggcaaat 1020

agaatactta aatctcgcac caaagcagtt cctggagcta cgcaggcaat tgcagagctg 1080

gataaggtat tggcatttaa taatttactc atctcattta agaatcatcc agatgtggat 1140

cgctttgctc gtggtgttgg tccaatttct ttagtgggtg gagagtatga tggtgacaga 1200

aaaatagagg acttgaaact cctttacagg gcatatgttt cagatgcttt gtctggtggg 1260

cgcatggaag atgataagct tgctgcacta aatcagttac gaaatatatt cggactgggt 1320

aaacgtgaag ccgaggccat ttcacttgat gttacgtcga aggtatatcg caaacgactt 1380

gcacaggctg ctgcagatgg tgagttagaa atggctgata gcaaagcagc cttccttcaa 1440

aatctatgtg atgaacttca ttttgatcca caaaaggcca gtgaacttca tgaagaaatt 1500

tatcggcaaa agcttcagag atgtgtagct gatggggagc tcaatgagga ggatgttgct 1560

gctttgttga ggatgcgtgt aatgctctgc ataccgcaac agattgttga agcagctcat 1620

tcagatatct gtggcagttt gtttgaaaag gttgtcaagg aggcaattgc atcaggggtt 1680

gatggatatg atgctgaaat ccagaaatca gtaagaaaag cagcacatgg cctgcgactt 1740

actagggagg ttgctatgtc tattgcaagc aaggcggtaa ggaagatttt tattaattac 1800

ataaaacgtg cacgggcagc tggaaatcgt accgagtctg cgaaagaact gaagaagatg 1860

atagccttca acaccttagt tgtaactaat ttggtggagg acattaaagg ggagtcaact 1920

gatatttcct ctgaagaacc tgtgaaagag gacattacac aaactgacga tgaagaatgg 1980

gaatcacttc agacactcaa gaaaataaga ccaaataaag aacttacgga aaagttggga 2040

aaacctggtc agacagaaat tactctgaaa gatgatcttc ctgaaaggga taggactgat 2100

ctttacaaga catacttact ttattgtcta actggtgaag tgacaagggt tccatttggt 2160

gctcagatca ctactaagaa ggatgattct gagtatcttc ttctaaatca gcttggtggg 2220

atcctgggat tgagtagtca agaaatagtg gaagtgcaca ggggtctagc tgagcaggct 2280

tttaggcaac aggctgaggt aattttagcc gatggacagt tgacaaaggc cagggtggag 2340

cagcttaata acctccagaa acaagtaggc ttacctcaag aatatgctca gaaaataatc 2400

aagagtataa ccactacaaa aatggcagct gccattgaaa ctgctgtaac tcaagggagg 2460

ctcaatatga agcagataag ggaacttaag gaagctgatg ttgatttaga cagtatggta 2520

tctgagaact tgagagagac cctcttcaaa aaaactgttg atgacatttt ctcatccggt 2580

actggagagt ttgacactga ggaagtatat gaaaaaatcc cgtcagatct caacattaac 2640

aaagagaagg cacgaggtgt tgttcatgag cttgcaaagg gtagactatc caactctctg 2700

attcaggctg tctctctact aagacagaga aatcagcagg gagtggtttc ttcactcaat 2760

gacttgctgg catgtgacaa agcagtaccc tcacagccag tttcatggga agtgccagag 2820

gagctttctg atctatacac catatacttg aagagtaatc caactcctga gaatttgtct 2880

cgtttgcaat atctgttggg tataaatgat tccacagctg ctgctcttag ggagattgga 2940

gatagattac tcaatactac tgcggaggaa gagaagtttg tattctag 2988

<210> 4

<211> 24

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 4

gattgcggcg gctggatacg gcct 24

<210> 5

<211> 24

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 5

aaacaggccg tatccagccg ccgc 24

<210> 6

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 6

atgaaccctt ccacactcac c 21

<210> 7

<211> 23

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 7

ctagaataca aacttctctt cct 23

<210> 8

<211> 7280

<212> DNA

<213> 人工序列(Artificial Sequence)

<400> 8

acgacgttat ctacaatcta caaagaggct ctatcatctt cttcctcttt agttttttcg 60

ttaaacccta taataacaac cactccactc ttctctactc tccgcaacct tacgacgtcg 120

tctcaaccat gaacccttcc acactcaccc cttcccacac ccaccgtcct ctcctcctac 180

cctctccctt ccacaccaga cggcgtcgtt ttaaagtctc cctccctcgc tgttcctcct 240

cctccgccgc ctcctccccc cctcctccgc cacctccgcc gccgcagcga ccgcccaagg 300

acctcaaggg aatcgatgtc ctcgtcgaca agctctcgcc gccggccagg ctcgccacct 360

ccgccgtcat cgtcgccggc gccgcggcgg ctggatacgg cctcggctcc cgcttcggcg 420

gaagccgcta cgctgcgctc ggtggagctg tcgccctcgg cgcggccgac ggtgctgcgg 480

cgtacgctct gaacgcggct gctccgcaag tcgccgccgt gaatttgcac aactacgtcg 540

ccgcgttcga cgacccttcg aagctgaaga aggaagagat tgaggccatt gcgtccaagt 600

tagtttttag tttcaatttc taataataat aactgttcat ttttttttct ttgttaattt 660

gagtgtttgg ttttgaaatg gttaattttt caggtatggt gtgagcaagc aagatgaggc 720

attcaaaact gagatttgtc atatatattc cgagtgagtg acaaattatt attattgttg 780

ttgttgtact ggtaacgagt tattatttgt tacttgcatt tgtgtagtga taatgtgtta 840

tgatgatttg gctgtttaat tttataattg aaagattgat ttgtggcata acgtgttgtt 900

caggtttgta tcttctgtgc ttcctcctgg tggtgaggaa cttaaaggtg atgaggttga 960

taggattgtc agcttcaaaa attctttggg aattgatgac ccggatgctg ctgctatgca 1020

catggaggta ttttttgtgt ttgttgtgta tatatataca cctgattacc agagtagatc 1080

gagtgtgtat gtttattatg aggagtgaaa gcagttccgt gtcttactgg tttggtgctt 1140

gttgactccc tagattggga ggaaattttt caggcaaagg ttggaagttg gggatcgtga 1200

tgctgatgtt gagcaacgtc gggtaagttt gttgtgttgt gatgttatct taactgattg 1260

gttgcaaatt tgcaatagca tcatgggttt tagatgaata atggatactt ggagcttgac 1320

cattgttaga gtctccagtg caggcatttc aaaagttgat atatgtgtca aatcttgtat 1380

ttggagatgc atcgtctttc cttctacctt ggaagcgtgt attcaaggtc accgattccc 1440

aggtatttat ttcttattgg ttaactggta tgcctagctt gtgcaacgtg atattaatat 1500

gcaattagga tgagagattt ggagcatagg tttatgacta tttataggca tttgctcaaa 1560

aagggaagac ctaatttgct gcacaaaatg cattttgcca tgtttgtgat cagtgacatg 1620

ctggcagcat ttcttaataa ttcttatatg ttctcattgt catttcttat cttagtgttt 1680

ttgttatttt tcatttgact agttaactat aatctattta ccagattgaa gtagctgtac 1740

gtgacaatgc gcagcgattg tttgcttcca agctgaaatc agttggcaga ggtaattgtg 1800

tagtcagtaa ttcctcttgg tccttgtttg ctcttactgt aacaaagttg atcaactaac 1860

tcacatttac tcctcgagaa gttgtcataa cttcatacta ctatattact ggagaaatga 1920

ttggaaatga taatcttgtt ttgtaattgt gcatgcattt cttattaaat aaagccttta 1980

aattttgctt gtactcatct agtatgctgg attttgttct gtaattctct agaattacta 2040

tcatccaaac taacaaattc tttctatagc aatttttaga agttttaaat caggtttatt 2100

tcctaagtgt taaccaatga tttcttgcca gatattgatg cagaaaaact tgttgcacta 2160

agaaaagaac aacaattatg tcgcctttct gatgaggttc attttcatta tttcttcttt 2220

cataactttc aacttctctg ggttaaatcc ctgtctttgt cttgccatca aatagttcct 2280

tggtatagta taatcttttt attaattaaa ggaaattttc attgaagctt gctgagaact 2340

tgtttagaga tcacacaagg aaattggttg aggaaaatat ttcagaggca aatagaatac 2400

ttaaatctcg caccaaagca gtgtaagatt catggttttt ctaagtattc ttttttttaa 2460

gtattaaatt ttgaattctg ggcatgtttt tcttgccatt tgtttcatta gtgttactat 2520

tcatttctta atttttacaa aaatagaatt tgccatatct gatcatctgt aagtttcagt 2580

cctggagcta cgcaggcaat tgcagagctg gataaggtat tggcatttaa taatttactc 2640

atctcattta agaatcatcc agatgtggat cgctttgctc gtggtgttgg tccaatttct 2700

ttagtgggta agtgtctgtg actaggtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgta 2760

taacatgttc ttttctcttt ttaatcttaa ttcttgaagt ttaccacaat tcatttttgg 2820

cataggtgga gagtatgatg gtgacagaaa aatagaggac ttgaaactcc tttacagggc 2880

atatgtttca gatgctttgt ctggtgggcg catggaagat gataaggtaa ataaccaaga 2940

caattggata tgaactgtga acttcttaag atgtgttttc tcaatattct aatatatttt 3000

gaagttgagt ttgggttagt gttttccaga tgtatgtagt gattgaatag tatattttct 3060

aaaaaaattg gagtattcag cagctattta tatcatattg gagtattcac ttcccatggg 3120

tataatagag attgaaacct agcagtttag ccttagttat tctttattgt ttcttatgtt 3180

ttatctttct ttttgatgta gtaccaattg ccaaatcatg attgaacagg ctgttcagtg 3240

catgctacca tatccatatt gctcttattc ttatttttaa tatttcagtt agggcagtat 3300

attattttca gtacaagggt atttgtttac tgtatcaagc ctaatgaatt ttcacttatt 3360

gaagcatttt ttagtgttat aaaacaatgt cctgtctcct tgcttaacac taacaactta 3420

aagcttttag gtcaggtgat ttgttgactg ttaagatgta ctagcatacg attccccttt 3480

gaatatagtt gcatatgttc tctaatcttc ctttaatgag tcaacattca gataagcaaa 3540

gacacaacag tgtctcgaaa accatgatat ccattactaa agagttctta ttgatataag 3600

atatgaaact tgatctgtca aaacacttat tttttaatat aaaataaaaa atatatttta 3660

aatatattta ttattcgttg cagcttgctg cactaaatca gttacgaaat atattcggac 3720

tgggtaaacg tgaagccgag gccatttcac ttgatgttac gtcgaaggta tatcgcaaac 3780

gacttgcaca ggctgctgca gatggtgagt tagaaatggc tgatagcaaa gcagccttcc 3840

ttcaaaatct atgtgatgaa cttcattttg atccacaaaa ggccagtgaa cttcatgaag 3900

gtaaaactcg tcatgtttac agttagttgg aagttgacta tatttgaggc ctttattatt 3960

gaacacttta tgtgctgtcg gtttttattt atgtgcaatt cttaaatctt aaggttgtaa 4020

aatgtaaact tgtaaaaccc tttctacatg atgtgacttt tttttatata tatatttagt 4080

tataacacaa tctaatgata gtttattatt tttaagaaat ttatcggcaa aagcttcaga 4140

gatgtgtagc tgatggggag ctcaatgagg aggatgttgc tgctttgttg aggatgcgtg 4200

taatgctctg cataccgcaa cagattgttg aagcagctca ttcagatatc tgtggcagtt 4260

tgtttgaaaa ggtagtcaac gttacttttt cctatctaat tgtttggtcc ttcataagtg 4320

tcaaattagt gaaattacat atttcttgct atgcagaagt gcggtccatc catgcactgt 4380

aacaaactga taatgataat atgaatacat ggatgaacac taaaattgct ttacatatga 4440

atatgggttc aagatgatta tggaactatg gttatcaaac tcattctact ttctatttga 4500

atattggtag catgagaagt tacagctgtc acattttaat gtgttgataa acctgataat 4560

gctttcattt actggtagtg tggagatgga gaccatgaat ctactgcatt tttttttatc 4620

ttacaggaaa gatacactgt tgtactctgt actctgcatg ctaggttttg tctagaattt 4680

ctaaaatgtg tatctattgg aattgttact ttcaactgct tgtaatttat ttgggctttg 4740

tcttttcaat ttgtagacct gtatctactt gctataaaat ctgtttattg gtttgattat 4800

cttatatcaa agttgtacaa gtctagattc tatagtgctt agttgatggc agcgatatgt 4860

gaagaaatgg gtctgtttta ttttgatata tcatgagtct tgtttcaatc tccatcaaag 4920

atgttcaaaa ttttgttgag ttattactaa ttcatgtttc tccttttata ggttgtcaag 4980

gaggcaattg catcaggggt tgatggatat gatgctgaaa tccagaaatc agtaagaaaa 5040

gcagcacatg gcctgcgact tactagggag gttgctatgt ctattgcaag caaggcggtg 5100

agtatcaagt tatcagaaac tcaatggtga tttataaaga tttgaatagg tttaaatttc 5160

actgaggttt tttggaatta taggtaagga agatttttat taattacata aaacgtgcac 5220

gggcagctgg aaatcgtacc gagtctgcga aagaactgaa gaagatgata gccttcaaca 5280

ccttagttgt aactaatttg gtggaggaca ttaaagggga gtcaactgat atttcctctg 5340

aagaacctgt gaaagaggac attacacaaa ctgacgatga agaatgggaa tcacttcaga 5400

cactcaagaa aataagacca aataaagaac ttacggaaaa gttgggaaaa cctggtcaga 5460

cagaaattac tctgaaagat gatcttcctg aaagggatag gactgatctt tacaagacat 5520

acttacttta ttgtctaact ggtgaagtga caagggttcc atttggtgct cagatcacta 5580

ctaagaagga tgattctgag tatcttcttc taaatcagct tggtgggatc ctgggattga 5640

gtagtcaaga aatagtggaa gtgcacaggg gtctagctga gcaggctttt aggcaacagg 5700

ctgaggtaat tttagccgat ggacagttga caaaggccag ggtggagcag cttaataacc 5760

tccagaaaca agtaggctta cctcaagaat atgctcagaa aataatcaag agtataacca 5820

ctacaaaaat ggcagctgcc attgaaactg ctgtaactca agggaggctc aatatgaagc 5880

agataaggga acttaaggaa gctgatgttg atttagacag tatggtatct gagaacttga 5940

gagagaccct cttcaaaaaa actgttgatg acattttctc atccggtact ggagagtttg 6000

acactgagga agtatatgaa aaaatcccgt cagatctcaa cattaacaaa gagaaggcac 6060

gaggtgttgt tcatgagctt gcaaagggta gactatccaa ctctctgatt caggctgtct 6120

ctctactaag acagagaaat cagcagggag tggtaagtaa tattcaaagc ttcatgttca 6180

cttagtacta gtttggattt atgctggatt atctaaaatt acactcaaat actagatcac 6240

gtgaagtaaa tgttcataaa ttttttgttt cactttaaga aattgattct gagttgaagc 6300

aattttaggt aactttttct gttgaataaa aaaggtttat gttaagtttt attattaact 6360

tgctttcatt aaaaaaaaac ctccaaacat aaatcttttc acttcaaaat caattatgca 6420

aaatctactt cattcataat caattttgca aatgtttgtc ccaacacact tagtgcgtgt 6480

ttgtacaagc attggcaaat tgattttaaa ttgaattgat tttagttaaa ttgatttgaa 6540

agtgatgtga tttatgttta agtacaacat tttagatcca tcgtaaaaat tactcaaagg 6600

tcctttaact tagagtcaat tttggactta gaatcaattc tgaattcttc tttaacagga 6660

aaccaaacac gaaaatttat caaaaagcca ttctccaatg tgaaacaaaa cttgcactta 6720

ttcaaacagc ttttttacgt ttgtttgtca tggataatca tgtgtgattg cttcttttct 6780

tttgtgcagg tttcttcact caatgacttg ctggcatgtg acaaagcagt accctcacag 6840

ccagtttcat gggaagtgcc agaggagctt tctgatctat acaccatata cttgaagagt 6900

aatccaactc ctgagaattt gtctcgtttg caatatctgt tgggtataaa tgattccaca 6960

gctgctgctc ttagggagat tggagataga ttactcaata ctactgcgga ggaagagaag 7020

tttgtattct agtcttgtga taaaatgttt ggcattttag gcacactaga gaggctgcaa 7080

gtccccttaa ttttgccatt ttttattttc cattttgttt aacggtagca gaaaaagaat 7140

atcacgatgt ataaactttg gaatagagat gagtgcagtg ctgaaatctt agataatatt 7200

ttgtttgaga cagttgaatt aatcaattgc tattttttat gaatgaaaat aagttttcat 7260

aatcgtggac aagttttatc 7280

<210> 9

<211> 995

<212> PRT

<213> 人工序列(Artificial Sequence)

<400> 9

Met Asn Pro Ser Thr Leu Thr Pro Ser His Thr His Arg Pro Leu Leu

1 5 10 15

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

20 25 30

Pro Arg Cys Ser Ser Ser Ser Ala Ala Ser Ser Pro Pro Pro Pro Pro

35 40 45

Pro Pro Pro Pro Gln Arg Pro Pro Lys Asp Leu Lys Gly Ile Asp Val

50 55 60

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

65 70 75 80

Ile Val Ala Gly Ala Ala Ala Ala Gly Tyr Gly Leu Gly Ser Arg Phe

85 90 95

Gly Gly Ser Arg Tyr Ala Ala Leu Gly Gly Ala Val Ala Leu Gly Ala

100 105 110

Ala Asp Gly Ala Ala Ala Tyr Ala Leu Asn Ala Ala Ala Pro Gln Val

115 120 125

Ala Ala Val Asn Leu His Asn Tyr Val Ala Ala Phe Asp Asp Pro Ser

130 135 140

Lys Leu Lys Lys Glu Glu Ile Glu Ala Ile Ala Ser Lys Tyr Gly Val

145 150 155 160

Ser Lys Gln Asp Glu Ala Phe Lys Thr Glu Ile Cys His Ile Tyr Ser

165 170 175

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

180 185 190

Asp Glu Val Asp Arg Ile Val Ser Phe Lys Asn Ser Leu Gly Ile Asp

195 200 205

Asp Pro Asp Ala Ala Ala Met His Met Glu Ile Gly Arg Lys Phe Phe

210 215 220

Arg Gln Arg Leu Glu Val Gly Asp Arg Asp Ala Asp Val Glu Gln Arg

225 230 235 240

Arg Ala Phe Gln Lys Leu Ile Tyr Val Ser Asn Leu Val Phe Gly Asp

245 250 255

Ala Ser Ser Phe Leu Leu Pro Trp Lys Arg Val Phe Lys Val Thr Asp

260 265 270

Ser Gln Ile Glu Val Ala Val Arg Asp Asn Ala Gln Arg Leu Phe Ala

275 280 285

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

290 295 300

Ala Leu Arg Lys Glu Gln Gln Leu Cys Arg Leu Ser Asp Glu Leu Ala

305 310 315 320

Glu Asn Leu Phe Arg Asp His Thr Arg Lys Leu Val Glu Glu Asn Ile

325 330 335

Ser Glu Ala Asn Arg Ile Leu Lys Ser Arg Thr Lys Ala Val Pro Gly

340 345 350

Ala Thr Gln Ala Ile Ala Glu Leu Asp Lys Val Leu Ala Phe Asn Asn

355 360 365

Leu Leu Ile Ser Phe Lys Asn His Pro Asp Val Asp Arg Phe Ala Arg

370 375 380

Gly Val Gly Pro Ile Ser Leu Val Gly Gly Glu Tyr Asp Gly Asp Arg

385 390 395 400

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

405 410 415

Leu Ser Gly Gly Arg Met Glu Asp Asp Lys Leu Ala Ala Leu Asn Gln

420 425 430

Leu Arg Asn Ile Phe Gly Leu Gly Lys Arg Glu Ala Glu Ala Ile Ser

435 440 445

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

450 455 460

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

465 470 475 480

Asn Leu Cys Asp Glu Leu His Phe Asp Pro Gln Lys Ala Ser Glu Leu

485 490 495

His Glu Glu Ile Tyr Arg Gln Lys Leu Gln Arg Cys Val Ala Asp Gly

500 505 510

Glu Leu Asn Glu Glu Asp Val Ala Ala Leu Leu Arg Met Arg Val Met

515 520 525

Leu Cys Ile Pro Gln Gln Ile Val Glu Ala Ala His Ser Asp Ile Cys

530 535 540

Gly Ser Leu Phe Glu Lys Val Val Lys Glu Ala Ile Ala Ser Gly Val

545 550 555 560

Asp Gly Tyr Asp Ala Glu Ile Gln Lys Ser Val Arg Lys Ala Ala His

565 570 575

Gly Leu Arg Leu Thr Arg Glu Val Ala Met Ser Ile Ala Ser Lys Ala

580 585 590

Val Arg Lys Ile Phe Ile Asn Tyr Ile Lys Arg Ala Arg Ala Ala Gly

595 600 605

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

610 615 620

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

625 630 635 640

Asp Ile Ser Ser Glu Glu Pro Val Lys Glu Asp Ile Thr Gln Thr Asp

645 650 655

Asp Glu Glu Trp Glu Ser Leu Gln Thr Leu Lys Lys Ile Arg Pro Asn

660 665 670

Lys Glu Leu Thr Glu Lys Leu Gly Lys Pro Gly Gln Thr Glu Ile Thr

675 680 685

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

690 695 700

Tyr Leu Leu Tyr Cys Leu Thr Gly Glu Val Thr Arg Val Pro Phe Gly

705 710 715 720

Ala Gln Ile Thr Thr Lys Lys Asp Asp Ser Glu Tyr Leu Leu Leu Asn

725 730 735

Gln Leu Gly Gly Ile Leu Gly Leu Ser Ser Gln Glu Ile Val Glu Val

740 745 750

His Arg Gly Leu Ala Glu Gln Ala Phe Arg Gln Gln Ala Glu Val Ile

755 760 765

Leu Ala Asp Gly Gln Leu Thr Lys Ala Arg Val Glu Gln Leu Asn Asn

770 775 780

Leu Gln Lys Gln Val Gly Leu Pro Gln Glu Tyr Ala Gln Lys Ile Ile

785 790 795 800

Lys Ser Ile Thr Thr Thr Lys Met Ala Ala Ala Ile Glu Thr Ala Val

805 810 815

Thr Gln Gly Arg Leu Asn Met Lys Gln Ile Arg Glu Leu Lys Glu Ala

820 825 830

Asp Val Asp Leu Asp Ser Met Val Ser Glu Asn Leu Arg Glu Thr Leu

835 840 845

Phe Lys Lys Thr Val Asp Asp Ile Phe Ser Ser Gly Thr Gly Glu Phe

850 855 860

Asp Thr Glu Glu Val Tyr Glu Lys Ile Pro Ser Asp Leu Asn Ile Asn

865 870 875 880

Lys Glu Lys Ala Arg Gly Val Val His Glu Leu Ala Lys Gly Arg Leu

885 890 895

Ser Asn Ser Leu Ile Gln Ala Val Ser Leu Leu Arg Gln Arg Asn Gln

900 905 910

Gln Gly Val Val Ser Ser Leu Asn Asp Leu Leu Ala Cys Asp Lys Ala

915 920 925

Val Pro Ser Gln Pro Val Ser Trp Glu Val Pro Glu Glu Leu Ser Asp

930 935 940

Leu Tyr Thr Ile Tyr Leu Lys Ser Asn Pro Thr Pro Glu Asn Leu Ser

945 950 955 960

Arg Leu Gln Tyr Leu Leu Gly Ile Asn Asp Ser Thr Ala Ala Ala Leu

965 970 975

Arg Glu Ile Gly Asp Arg Leu Leu Asn Thr Thr Ala Glu Glu Glu Lys

980 985 990

Phe Val Phe

995