本發(fā)明涉及致病基因，尤其是涉及一種嚴(yán)重弱精子癥新致病基因及其應(yīng)用。

背景技術(shù)：

嚴(yán)重弱精子癥是一種較常見的導(dǎo)致男性不育的畸形精子癥，是一種與遺傳、生活方式等因素有關(guān)的疾病，其主要臨床特征是精子運(yùn)動(dòng)能力的嚴(yán)重喪失。國(guó)外早期的分子方面發(fā)病機(jī)制研究發(fā)現(xiàn)akap3、akap4、slc26a8、dnah1、catsper2、galntl5等基因發(fā)生了突變(1.takasakin,tachibanak,ogasawaras,matsuzakih,hagiudaj,ishikawah,mochidak,inouek,ogonukin,oguraa,nocet,itoc,toshimorikandnarimatsuh.aheterozygousmutationofgalntl5affectsmaleinfertilitywithimpairmentofspermmotility.procnatlacadsciusa.2014；111(3):1120-1125；2.diramit,rodeb,jollivetm,dasilvan,escalierd,gaitchn,norezc,tufferyp,wolfjp,becqf,raypf,dμliouste,gacong,bienvenutandtourea.missensemutationsinslc26a8,encodingasperm-specificactivatorofcftr,areassociatedwithhumanasthenozoospermia.amjhumgenet.2013；92(5):760-766.)，并導(dǎo)致精子尾部形態(tài)及運(yùn)動(dòng)異常。

目前嚴(yán)重弱精子癥的基因突變譜尚未完全發(fā)現(xiàn)，基因型和表現(xiàn)型的關(guān)系也不明確。目前單基因疾病的研究開始大量采用全外顯子組測(cè)序(whole-exomesequencing)及全基因組測(cè)序(whole-genomesequencing)的方法，這兩種方法被成功的應(yīng)用于發(fā)現(xiàn)稀有單基因疾病的致病基因。全外顯子組測(cè)序及全基因組測(cè)序技術(shù)已被證明為降低稀有單基因疾病候選基因甚至發(fā)現(xiàn)其致病基因的有力、有效手段。僅通過對(duì)幾個(gè)很少的個(gè)體(包括患者及正常對(duì)照)的全外顯子組或基因組進(jìn)行測(cè)序來篩選與疾病相關(guān)的變異，其成功率大為提升。

因此本領(lǐng)域?qū)?yán)重弱精子癥的研究尚不清晰，對(duì)造成該疾病的原因更不明了，因此本領(lǐng)域迫切需要對(duì)嚴(yán)重弱精子癥的致病機(jī)理進(jìn)行研究，找到新致病基因及突變位點(diǎn)。

技術(shù)實(shí)現(xiàn)要素：

本發(fā)明的第一目的在于提供通過外顯子組測(cè)序的方法確定嚴(yán)重弱精子癥的新致病基因。

本發(fā)明的第二目的在于提供嚴(yán)重弱精子癥的生物標(biāo)記物。

本發(fā)明的第三目的在于提供突變的spag17基因。

本發(fā)明的第四目的在于提供突變的spag17蛋白。

本發(fā)明的第五目的在于提供一種檢測(cè)嚴(yán)重弱精子癥的方法。

本發(fā)明的第六目的在于提供通過pcr檢測(cè)spag17基因或spag17蛋白突變中使用的引物對(duì)。

本發(fā)明的第七目的在于提供與突變spag17基因互補(bǔ)的核酸探針。

本發(fā)明的第八目的在于提供檢測(cè)突變spag17基因或spag17蛋白的試劑盒。

本發(fā)明的第九目的在于提供檢測(cè)突變spag17基因的試劑盒。

本發(fā)明的第十目的在于提供一種嚴(yán)重弱精子癥新致病基因的應(yīng)用。

所述通過外顯子組測(cè)序的方法確定嚴(yán)重弱精子癥的新致病基因。

所述嚴(yán)重弱精子癥的生物標(biāo)記物，即突變的spag17基因或spag17蛋白，所述生物標(biāo)記物是具有選自如下的突變的spag17基因或spag17蛋白：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)。

所述突變的spag17基因?yàn)閟eqidno:1的序列中具有以下突變：

在外顯子30中錯(cuò)義突變(c.4343g>a)。

所述突變的spag17蛋白為seqidno:2的序列中具有以下突變：

在外顯子30中錯(cuò)義突變(p.r1448q)。

所述一種檢測(cè)嚴(yán)重弱精子癥的方法，包括檢測(cè)受試者的spag17基因或spag17蛋白中是否存在突變位點(diǎn)，若有突變位點(diǎn)，則所述受試者被鑒定為患有嚴(yán)重弱精子癥或易患嚴(yán)重弱精子癥，所述突變位點(diǎn)選自如下一種：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)。

所述spag17蛋白為seqidno:2的序列表示。

所述spag17基因?yàn)閟eqidno:1的序列表示。

所述檢測(cè)嚴(yán)重弱精子癥的方法包括如下至少一組引物擴(kuò)增的步驟：

seqidno:3和seqidno:4。

所述檢測(cè)嚴(yán)重弱精子癥的方法中檢測(cè)突變位點(diǎn)通過選自如下進(jìn)行：測(cè)序、電泳、核酸雜交、原位雜交、pcr、逆轉(zhuǎn)錄酶鏈反應(yīng)和變性高效液相色譜。

所述通過pcr檢測(cè)spag17基因或spag17蛋白突變中使用的引物對(duì)，所述突變是選自如下一種：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)，

其中所述引物對(duì)分別基于選自如下的位置前后設(shè)計(jì)，使得擴(kuò)增該位置(編號(hào)基于spag17的cdna序列)：4343。

所述與突變spag17基因互補(bǔ)的核酸探針，所述突變是選自如下一種：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)，

所述探針與突變spag17基因的互補(bǔ)區(qū)包括選自如下的位置(編號(hào)基于spag17的cdna序列)：4343。

所述檢測(cè)突變spag17基因或spag17蛋白的試劑盒，包含一組或多組引物對(duì)，其中所述突變是選自如下一種：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)，

其中所述引物對(duì)分別基于選自如下的位置前后設(shè)計(jì)，使得其擴(kuò)增產(chǎn)物涵蓋該位置(編號(hào)基于spag17的cdna序列)：4343。

所述檢測(cè)突變spag17基因或spag17蛋白的試劑盒包含選自如下的至少一組引物：

seqidno:3和seqidno:4。

所述檢測(cè)突變spag17基因的試劑盒，包含一個(gè)或多個(gè)核酸探針，所述突變是選自如下一種：

在外顯子30中錯(cuò)義突變(c.4343g>a；p.r1448q)，

所述探針與突變spag17基因上包含選自如下的位置的區(qū)域互補(bǔ)(編號(hào)基于spag17的cdna序列)：4343。

所述一種嚴(yán)重弱精子癥新致病基因的應(yīng)用。

本發(fā)明將為嚴(yán)重弱精子癥的發(fā)病機(jī)制研究奠定重要基礎(chǔ)，有可能為嚴(yán)重弱精子癥的患者治療提供全新的理論依據(jù)。

具體實(shí)施方式

在本發(fā)明中，采用本領(lǐng)域通用表示法表示突變。例如，突變(c.4343g>a；p.r1448q)中，c表示cdna，p表示蛋白質(zhì)，dna水平的突變對(duì)應(yīng)蛋白質(zhì)水平的突變。

野生型spag17基因的cdna序列如seqidno:1所示。

seqidno:2：野生型spag17蛋白的氨基酸序列。

提及基因序列，本領(lǐng)域技術(shù)人員應(yīng)當(dāng)理解，實(shí)際包括互補(bǔ)雙鏈的任意一條，或者兩條。雖然多數(shù)情況下至給出了一條鏈，但實(shí)際上也公開了與之互補(bǔ)的另一條鏈。例如提及spag17基因的cdna序列，實(shí)際上包括該序列以及其互補(bǔ)序列。例如，提及seqidno:1，實(shí)際包括其互補(bǔ)序列。本領(lǐng)域技術(shù)人員還可以理解，利用一條鏈可以檢測(cè)另一條鏈，反之亦然。

本申請(qǐng)中的基因序列包括dna形式或rna形式，公開其中一種，意味著另一種也被公開。例如提及spag17基因的cdna序列，實(shí)際也包括相應(yīng)的rna序列。

實(shí)施例1確定嚴(yán)重弱精子癥的致病基因。

本發(fā)明收集1個(gè)近親婚配家系中的兩位雙胞胎患有嚴(yán)重弱精子癥的病例，兩位患者均表現(xiàn)為不育。先證者29歲，父母為三代內(nèi)姑表親結(jié)婚，生育3男1女，哥哥生育1子，妹妹生育兩胎，雙胞胎弟弟不育。先證者外生殖器發(fā)育正常，雙側(cè)睪丸大小正常，雙側(cè)精索靜脈未觸及異常。外周血染色體核型分析正常，y染色體微缺失檢測(cè)未發(fā)現(xiàn)異常。無支氣管擴(kuò)張等上呼吸道疾病，無內(nèi)臟轉(zhuǎn)位，無鼻竇炎病史。在我院三次精液檢查平均值為精液量2.8ml，精子密度27.5百萬/ml，前向運(yùn)動(dòng)精子占比1.8％，非前向運(yùn)動(dòng)精子1.6％。精子形態(tài)改良巴氏染色讀取顯示正常形態(tài)精子4.5％，精子頂體酶28miu/ml，精漿生化果糖、中性糖苷酶活性及精漿鋅均未見異常，根據(jù)以上檢查結(jié)果提示患者為嚴(yán)重弱精子癥。

本發(fā)明對(duì)先癥者進(jìn)行了全外顯子組測(cè)序，具體步驟如下：

樣品制備：采集所述患者及其父母家人外周血，利用試劑盒抽提外周血白細(xì)胞中的基因組dna(qiaampdnaminikit51304.qiagen,usa)，利用nanodrop2000測(cè)量dna的濃度及純度(thermoscientific,usa)，所得的每個(gè)標(biāo)本基因組dna的od260/od280均位于1.7～2.0之間，濃度不少于100ng/μl，總量不少于30μl。

然后，對(duì)上述1個(gè)樣本的外顯子組序列進(jìn)行了測(cè)序。測(cè)序平臺(tái)為illuminahiseq2000，依照illumina標(biāo)準(zhǔn)建庫說明書(參見http://www.illumina.com/)進(jìn)行測(cè)序，簡(jiǎn)述如下：

1)使用illumina公司試劑盒truseqdnalibraryprepkit制作dna文庫。在外顯子捕獲過程中使用agilent公司試劑盒sureselecthumanallexonv5；

2)使用illumina公司hiseq2000測(cè)序平臺(tái)對(duì)捕獲的外顯子區(qū)域進(jìn)行平行測(cè)序讀取長(zhǎng)度為90bp，每個(gè)樣本的平均測(cè)序深度最少為100；

3)使用burrows-wheeleraligner(http://bio-bwa.sourceforge.net/)將測(cè)序結(jié)果比對(duì)到人參考基因組中(hg19)。使用samtools將重復(fù)讀取結(jié)果去除，通過gatk(https://www.broadinstitute.org/gatk/)進(jìn)行調(diào)整和校準(zhǔn)。然后使用samtools(http://www.samtools.sourceforge.net/)挖掘出snvs及indels，并用annovar(http://www.openbioinformatics.org/annovar/)命名。

將等位基因頻率大于1％的突變位點(diǎn)剔除(參考數(shù)據(jù)庫包括dbsnp，1000genomes，exomeaggregationconsortium(exac))。

發(fā)現(xiàn)先癥者在基因spag17上具有純合突變：通過家系中2名患者和4位正常人snp篩查共分離，這些位點(diǎn)可能是致病位點(diǎn)。

結(jié)果顯示于表3。

因此本發(fā)明認(rèn)為spag17基因極有可能為嚴(yán)重弱精子癥的致病基因。

實(shí)施例2在其他病例中確認(rèn)上述嚴(yán)重弱精子癥的致病基因。

本發(fā)明納入所有嚴(yán)重弱精子癥患者，對(duì)spag17基因?yàn)閲?yán)重弱精子癥的致病基因進(jìn)行驗(yàn)證。驗(yàn)證中利用sanger法測(cè)序驗(yàn)證spag17基因的突變，其中考慮了家系內(nèi)等樣本驗(yàn)證情況。

分別對(duì)2名患者、4名家系內(nèi)正常人(即其中2名患者的父母及哥哥姐姐，他們均未發(fā)病)基因進(jìn)行檢測(cè)。

具體方法步驟如下：

1.dna提?。?/p>

分別提取2名患者、4名家系內(nèi)正常人的外周靜脈血，按照實(shí)施例1的方法提取基因組dna和測(cè)定dna含量。

2.引物設(shè)計(jì)及pcr反應(yīng)

引物設(shè)計(jì)參考人類基因組序列數(shù)據(jù)庫hg19，引物序列參見表1。

表1

pcr擴(kuò)增體系參見表2。

表2

pcr反應(yīng)條件：

94℃變性5min，然后進(jìn)入每個(gè)循環(huán)，即94℃變性30s，退火(通常為55℃，根據(jù)不同引物的退火溫度設(shè)定)30s，72℃延伸1min，總共設(shè)定30個(gè)循環(huán)。所有循環(huán)結(jié)束，最終72℃延伸10min，并將pcr產(chǎn)物保存在4或-20℃。

3)將步驟2)中獲得的獲自2名患者、4名家系內(nèi)正常人(即該2例患者的父母，他們均未發(fā)病)的pcr擴(kuò)征產(chǎn)物直接進(jìn)行dna測(cè)序，方法同實(shí)施例1。

本發(fā)明收集到的家系內(nèi)2例嚴(yán)重弱精子癥病例均在spag17基因中檢測(cè)到有意義的突變位點(diǎn)。同時(shí)所發(fā)現(xiàn)的突變位點(diǎn)在家族正常人中均未能檢測(cè)到。這些突變位點(diǎn)在exac數(shù)據(jù)庫中的頻率極低或無頻率(見表3)，提示所檢測(cè)到的位點(diǎn)很可能并非snp。因此認(rèn)為spag17基因?yàn)閲?yán)重弱精子癥的致病基因。

表3

序列表說明

seqidno:1：野生型spag17基因的cdna序列

atggcacccaagaaggagaaaggaggaactgtgaacaccagttctaagatatgggaaccctcgctcatagctgcacagttcaatcagaacgattggcaggcctccattgcttttgtggttgggaaccagattgaagatgatcttctcatccaagcccttaccgtggctgtccaggtccctcagcgtaaactcttcagtatggtgtcgtggcaagacattctccagcagattaatgaaataaatacacttgttggatctgcttcatctaaaaaggcaaaaaaacctgtaggtggtaatgctcctttatattatgaggtgttaacggcagcaaaagcaattatggatagtggagagaaattaaccttaccactgatagggaaactcttgaaatttcaacttctccagattaaatttaaggaccaacagcgacgggaaaatgaaaagaaggtaatagaagacaaacctaagttagaaaaggataaagggaaagcaaaatctcccaaggagaaaaaggctccaagtgccaagcctgccaaaggaaagggaaaggatcagcctgaggcaaatgcaccagtgaaaaagaccacccagttaaagcggagaggagaagacgaccacaccaatcgttacattgacgatgagccagatgatggtgcccaacattacattatagttgtgggctttaacaatcctcagctattagcaattatggctgagcttggcattcctataaccagcgtgattaaaatatcttcagagaattatgaacctctgcagacacacctggcagcagttaaccagcagcaggaagttcttcttcagtcagaagatctagaagcagaaaaattgaagaaagaaaatgccataaaagagcttaaaactttctggaagtacttggaaccagtcctgaataatgagaaacctgaaacaaatctctttgatgttgctcgacttgagtacatggtcaaagcagctgattttccttctgactggtcagatggtgagatgatgctgaaattgggcactgatatttttgaaaatattgcctgcttgatgtatgacatcctggattggaaaaggcagcaccagcactatttggaaagcatgcagcttattaatgttccacaagtggttaatgagaaacctgtattagaagccatgccaacttcagaggctccacaacctgctgtaccagctcctggaaagaagaaagcacagtatgaagaaccgcaagctccaccaccagtgacttcagtcatcacaactgaagtagacatgagatattacaattatttgctgaatccaattcgagaggaattcatttctgtgcccctgatactgcattgtatgctggaacaggttgttgcaactgaagaagatctcgtcccacccagtctgcgggagccatcccccagagcagacgggctagaccacagaatcgcagctcacattgtgtcccttctgccctcactctgtctctcagagagggagaaaaagaatcttcatgacatatttttatctgaagaagaaaatgaaagcaaagcagtgcccaaaggccccctcctactgaactatcatgatgcacacgcccacaagaagtacgcactacaggaccaaaagaattttgatccagttcaaattgagcaggagatgcagtccaagttgccactgtgggaatttcttcaattccctctacccccaccatggaacaacactaaacgtctagctacaattcatgagcttatgcacttttgtacgagtgatgtcttgagctggaatgaagtagaacgagccttcaaggtgtttacttttgagagcctgaagctctctgaggttgatgaaaaagggaaactgaaaccttctgggatgatgtgtgggtcagattctgaaatgttcaacataccgtgggacaaccctgccagatttgctaaacagataaggcagcaatatgtcatgaaaatgaatactcaagaggccaagcagaaagcagatattaaaatcaaagacagaacactatttgtggatcagaatttgtcaatgtctgtgcaagataatgaaagcaaccgagaaccttcagatcctagtcagtgtgatgctaacaatatgaagcattctgacttgaataatctcaaactctcagtccctgataatagacagctgttagagcaggagagcatcatgaaggctcagccccaacatgagtctctggagcagaccacaaacaatgagatcaaagatgatgcagtcacaaaggctgattctcatgaaaagaaacccaagaagatgatggtggaagcagatttagaggacataaagaaaacacagcagcgcagtctaatggactggagttttactgaacattttaaaccgaaagtactgcttcaggtccttcaagaagcccataagcaatataggtgtgttgattcttactaccacacccaagacaactctttacttttagtctttcacaatccaatgaatagacaacgtttgcattgtgaatattggaacattgctctccactccaatgttggattcaggaattatttggaacttgttgcaaaatctattcaagattggattacaaaagaagaagctatatatcaggaatctaaaatgaatgagaaaatcatcaggaccagagctgagctggaattgaaatcttctgctaatgccaaacttacttctgctagcaaaattttttccattaaagaatctaaaagtaacaaaggaatcagcaaaacagagatatcagatcaagaaaaagaaaaagagaaggaaaagattcctttcattttagaaggctctctcaaggcatggaaagaagagcaacatcgattagcagaagaggagcgcttaagggaagaaaagaaagcagagaagaagggtaaagaagctggtaaaaagaaaggcaaggataacgcagagaaagaggatagtaggtctttgaagaaaaaatcaccttacaaggagaaatctaaagaagaacaagtcaagatccaagaagtaacagaagagtccccccaccaaccagaacctaagataacttacccgtttcacggatacaatatgggaaatatacccactcaaatctcagggtcaaattactacctgtatccttctgatggggggcagattgaagtggaaaagacaatgtttgaaaaaggcccaacttttatcaaagtgagagtggtaaaggacaaccacaattttatgattcatttaaatgaccctaaggaaattgtgaaaaaggaagagaaaggggattattatttagaagaggaagaagaaggagatgaggaacaaagtcttgaaacggaagtatcagatgcaaagaataaagctttcagcaagtttggatctttttctgccaccttagaaaatggaatctgcctctcgataagttactatggatcaaatggaatggcaccagaagataaggatcctgatttagaaacaatattgaatatcccttcagcactcactccaacagtggttcc

tgttatagtgaccgttcctcaaagcaaagctaaagggaaaataaaaggcaaagaaaaacccaaagaatcccttaaagaagaagaacacccaaaagaagaagagaaaaaggaagaagaagtagaaccagaacctgttttacaagagactttggatgttcccaccttccagagcctaaatgtgtcttgccccagtgggctcctgttgactttcattggacaagaatctacaggtcaatatgttatagatgaggaacccacctgggacatcatggtccgtcagagctacccccagagggtgaagcactatgagttctataaaacggtgatgccacccgcagagcaggaggcttcaagggttatcaccagtcaaggcactgttgtcaaatatatgttggatggatccacacagattctctttgcagatggtgctgtgagcaggagtcccaattcaggtcttatttgtcctccttctgaaatgccagcaacgcctcacagtggagatttgatggactctatttctcagcagaaatcagaaacgataccatctgagattaccaacacaaagaaaggaaaaagtcacaaaagtcagtcatcaatggcccataagggtgaaatccatgaccctcctccagaggcagttcaaactgtaactcctgtggaggttcacataggcacctggtttacaaccacacctgaaggaaatcggatcggcaccaaaggattagaaagaatagcagacttgaccccattgttatcctttcaggccacagatcctgtcaatggaacggttatgacaactcgagaagacaaagttgtcatagttgaaaggaaagatggtactcggatagtggatcatgctgatggtaccagaatcacaaccttttatcaagtttatgaagatcaaattattctgccagatgatcaagaaacaaccgagggtcctcggactgtcaccaggcaggtgaagtgtatgcgggtagaaagctcacgctatgccactgttatcgccaactgtgaggacagtagctgctgtgccacctttggagatggaacaactattattgcaaagccacagggaacataccaggtgttacctccaaacacaggctctctttatattgacaaggattgttcagctgtgtactgccatgagtcaagcagtaatatatactatccttttcaaaagcgtgagcagctgcgagctggcaggtacatcatgaggcatacttcagaggttatctgtgaggttctggatcctgagggaaacacttttcaggtcatggctgatggtagcatatcaactatattacctgaaaaaaaattggaagatgatttaaatgagaaaactgagggctatgatagtctgtcctctatgcaccttgaaaagaatcatcagcaaatctatggtgaacatgtccccaggttttttgttatgtatgctgatggatcaggaatggaacttcttcgagacagtgacatagaagaatatctatctttggcatataaagaatcaaatactgttgttctccaagagccagtgcaggaacagccaggcaccctaaccatcacagtccttcgccctttccatgaagcatcaccatggcaagtaaaaaaggaagatacaattgtccctcctaatctccggtcaaggtcatgggaaacatttccctcagttgagaaaaaaactccaggacctccgtttggtactcagatttggaaaggcctttgcattgagtccaaacagctagtgagtgccccgggtgccatactcaagagccccagtgtgctacagatgcgccaattcattcagcatgaggtcataaagaatgaggtgaaactgaggctgcaggtttcccttaaggattacataaactatattctaaagaaagaagatgagctgcaggaaatgatggttaaagattccagaactgaggaggagagaggcaatgctgctgatctcctcaagctggttatgtctttccctaaaatggaggaaactacaaaaagtcatgttactgaagttgcagctcacctaactgatttattcaagcagtctttggctacgcctccaaaatgcccaccagacacatttggtaaagatttctttgaaaagacatggagacacacagcatcctcaaaacgctggaaagaaaagatagacaaaacgaggaaggaaattgagacaacacagaattacctaatggatattaagaaccgcataataccacccttttttaaatctgaattgaaccagttatatcagtctcagtataatcacctggacagtctttccaaaaaactgccttcttttacaaagaaaaatgaagatgcaaacgaaacagctgttcaagatacatctgatcttaatctagatttcaagccacataaggtttcagaacagaaatcctcaagtgtgcctagtcttccaaaaccagagatttctgcagataagaaggatttcactgctcagaaccaaactgaaaatttaacaaaatctcctgaagaagcagaatcttatgagcccgtgaaaattccaacccagtccttgctgcaggatgttgcgggacaaacaagaaaagaaaaagtgaagttgcctcattatttgctgagttccaagcctaagtctcaacctcttgcaaaggtgcaagattctgttggaggaaaagtgaacacatcctctgttgcatctgctgccattaataatgcaaagtcatccctttttgggttccatcttctcccatcatcagtcaagtttggagtgcttaaggaaggacatacctatgccacagttgtaaagctcaagaatgttggagtggacttctgcaggtttaaagtaaagcagcccccacccagcacaggactgaaagtgacttacaaacctggacctgtggcagctggtatgcagacagaactgaatatagagttatttgccacagctgttggagaggatggggccaagggatcagcacacatctctcacaatatcgagattatgacagagcatgaggttctgttcctacctgtggaagcaactgttttaacaagcagcaattatgataaacgaccaaaagactttccccagggaaaagaaaatccaatggtccagagaacttctacaatttattcctccacacttggagtcttcatgtctcgtaaagtttctccacattag

seqidno:2：野生型spag17蛋白的氨基酸序列

mapkkekggtvntsskiwepsliaaqfnqndwqasiafvvgnqieddlliqaltvavqvpqrklfsmvswqdilqqineintlvgsasskkakkpvggnaplyyevltaakaimdsgekltlpligkllkfqllqikfkdqqrrenekkviedkpklekdkgkakspkekkapsakpakgkgkdqpeanapvkkttqlkrrgeddhtnryiddepddgaqhyiivvgfnnpqllaimaelgipitsvikissenyeplqthlaavnqqqevllqsedleaeklkkenaikelktfwkylepvlnnekpetnlfdvarleymvkaadfpsdwsdgemmlklgtdifeniaclmydildwkrqhqhylesmqlinvpqvvnekpvleamptseapqpavpapgkkkaqyeepqapppvtsvittevdmryynyllnpireefisvplilhcmleqvvateedlvppslrepspradgldhriaahivsllpslclserekknlhdiflseeeneskavpkgplllnyhdahahkkyalqdqknfdpvqieqemqsklplweflqfplpppwnntkrlatihelmhfctsdvlswneverafkvftfeslklsevdekgklkpsgmmcgsdsemfnipwdnparfakqirqqyvmkmntqeakqkadikikdrtlfvdqnlsmsvqdnesnrepsdpsqcdannmkhsdlnnlklsvpdnrqlleqesimkaqpqhesleqttnneikddavtkadshekkpkkmmveadledikktqqrslmdwsftehfkpkvllqvlqeahkqyrcvdsyyhtqdnslllvfhnpmnrqrlhceywnialhsnvgfrnylelvaksiqdwitkeeaiyqeskmnekiirtraelelkssanakltsaskifsikesksnkgiskteisdqekekekekipfilegslkawkeeqhrlaeeerlreekkaekkgkeagkkkgkdnaekedsrslkkkspykekskeeqvkiqevteesphqpepkitypfhgynmgniptqisgsnyylypsdggqievektmfekgptfikvrvvkdnhnfmihlndpkeivkkeekgdyyleeeeegdeeqsletevsdaknkafskfgsfsatlengiclsisyygsngmapedkdpdletilnipsaltptvvpvivtvpqskakgkikgkekpkeslkeeehpkeeekkeeevepepvlqetldvptfqslnvscpsgllltfigqestgqyvideeptwdimvrqsypqrvkhyefyktvmppaeqeasrvitsqgtvvkymldgstqilfadgavsrspnsglicppsempatphsgdlmdsisqqksetipseitntkkgkshksqssmahkgeihdpppeavqtvtpvevhigtwftttpegnrigtkgleriadltpllsfqatdpvngtvmttredkvviverkdgtrivdhadgtrittfyqvyedqiilpddqettegprtvtrqvkcmrvessryatviancedssccatfgdgttiiakpqgtyqvlppntgslyidkdcsavychesssniyypfqkreqlragryimrhtsevicevldpegntfqvmadgsistilpekkleddlnektegydslssmhleknhqqiygehvprffvmyadgsgmellrdsdieeylslaykesntvvlqepvqeqpgtltitvlrpfheaspwqvkkedtivppnlrsrswetfpsvekktpgppfgtqiwkglcieskqlvsapgailkspsvlqmrqfiqheviknevklrlqvslkdyinyilkkedelqemmvkdsrteeergnaadllklvmsfpkmeettkshvtevaahltdlfkqslatppkcppdtfgkdffektwrhtasskrwkekidktrkeiettqnylmdiknriippffkselnqlyqsqynhldslskklpsftkknedanetavqdtsdlnldfkphkvseqksssvpslpkpeisadkkdftaqnqtenltkspeeaesyepvkiptqsllqdvagqtrkekvklphyllsskpksqplakvqdsvggkvntssvasaainnaksslfgfhllpssvkfgvlkeghtyatvvklknvgvdfcrfkvkqpppstglkvtykpgpvaagmqtelnielfatavgedgakgsahishnieimtehevlflpveatvltssnydkrpkdfpqgkenpmvqrtstiysstlgvfmsrkvsph。

序列表

<110>廈門市婦幼保健院（廈門市計(jì)劃生育服務(wù)中心），首都醫(yī)科大學(xué)附屬北京婦產(chǎn)醫(yī)院，煙臺(tái)毓璜頂醫(yī)院

<120>一種嚴(yán)重弱精子癥新致病基因及其應(yīng)用

<130>2017

<160>4

<170>patentinversion3.3

<210>1

<211>6672

<212>dna

<213>homosapiens

<400>1

atggcacccaagaaggagaaaggaggaactgtgaacaccagttctaagatatgggaaccc60

tcgctcatagctgcacagttcaatcagaacgattggcaggcctccattgcttttgtggtt120

gggaaccagattgaagatgatcttctcatccaagcccttaccgtggctgtccaggtccct180

cagcgtaaactcttcagtatggtgtcgtggcaagacattctccagcagattaatgaaata240

aatacacttgttggatctgcttcatctaaaaaggcaaaaaaacctgtaggtggtaatgct300

cctttatattatgaggtgttaacggcagcaaaagcaattatggatagtggagagaaatta360

accttaccactgatagggaaactcttgaaatttcaacttctccagattaaatttaaggac420

caacagcgacgggaaaatgaaaagaaggtaatagaagacaaacctaagttagaaaaggat480

aaagggaaagcaaaatctcccaaggagaaaaaggctccaagtgccaagcctgccaaagga540

aagggaaaggatcagcctgaggcaaatgcaccagtgaaaaagaccacccagttaaagcgg600

agaggagaagacgaccacaccaatcgttacattgacgatgagccagatgatggtgcccaa660

cattacattatagttgtgggctttaacaatcctcagctattagcaattatggctgagctt720

ggcattcctataaccagcgtgattaaaatatcttcagagaattatgaacctctgcagaca780

cacctggcagcagttaaccagcagcaggaagttcttcttcagtcagaagatctagaagca840

gaaaaattgaagaaagaaaatgccataaaagagcttaaaactttctggaagtacttggaa900

ccagtcctgaataatgagaaacctgaaacaaatctctttgatgttgctcgacttgagtac960

atggtcaaagcagctgattttccttctgactggtcagatggtgagatgatgctgaaattg1020

ggcactgatatttttgaaaatattgcctgcttgatgtatgacatcctggattggaaaagg1080

cagcaccagcactatttggaaagcatgcagcttattaatgttccacaagtggttaatgag1140

aaacctgtattagaagccatgccaacttcagaggctccacaacctgctgtaccagctcct1200

ggaaagaagaaagcacagtatgaagaaccgcaagctccaccaccagtgacttcagtcatc1260

acaactgaagtagacatgagatattacaattatttgctgaatccaattcgagaggaattc1320

atttctgtgcccctgatactgcattgtatgctggaacaggttgttgcaactgaagaagat1380

ctcgtcccacccagtctgcgggagccatcccccagagcagacgggctagaccacagaatc1440

gcagctcacattgtgtcccttctgccctcactctgtctctcagagagggagaaaaagaat1500

cttcatgacatatttttatctgaagaagaaaatgaaagcaaagcagtgcccaaaggcccc1560

ctcctactgaactatcatgatgcacacgcccacaagaagtacgcactacaggaccaaaag1620

aattttgatccagttcaaattgagcaggagatgcagtccaagttgccactgtgggaattt1680

cttcaattccctctacccccaccatggaacaacactaaacgtctagctacaattcatgag1740

cttatgcacttttgtacgagtgatgtcttgagctggaatgaagtagaacgagccttcaag1800

gtgtttacttttgagagcctgaagctctctgaggttgatgaaaaagggaaactgaaacct1860

tctgggatgatgtgtgggtcagattctgaaatgttcaacataccgtgggacaaccctgcc1920

agatttgctaaacagataaggcagcaatatgtcatgaaaatgaatactcaagaggccaag1980

cagaaagcagatattaaaatcaaagacagaacactatttgtggatcagaatttgtcaatg2040

tctgtgcaagataatgaaagcaaccgagaaccttcagatcctagtcagtgtgatgctaac2100

aatatgaagcattctgacttgaataatctcaaactctcagtccctgataatagacagctg2160

ttagagcaggagagcatcatgaaggctcagccccaacatgagtctctggagcagaccaca2220

aacaatgagatcaaagatgatgcagtcacaaaggctgattctcatgaaaagaaacccaag2280

aagatgatggtggaagcagatttagaggacataaagaaaacacagcagcgcagtctaatg2340

gactggagttttactgaacattttaaaccgaaagtactgcttcaggtccttcaagaagcc2400

cataagcaatataggtgtgttgattcttactaccacacccaagacaactctttactttta2460

gtctttcacaatccaatgaatagacaacgtttgcattgtgaatattggaacattgctctc2520

cactccaatgttggattcaggaattatttggaacttgttgcaaaatctattcaagattgg2580

attacaaaagaagaagctatatatcaggaatctaaaatgaatgagaaaatcatcaggacc2640

agagctgagctggaattgaaatcttctgctaatgccaaacttacttctgctagcaaaatt2700

ttttccattaaagaatctaaaagtaacaaaggaatcagcaaaacagagatatcagatcaa2760

gaaaaagaaaaagagaaggaaaagattcctttcattttagaaggctctctcaaggcatgg2820

aaagaagagcaacatcgattagcagaagaggagcgcttaagggaagaaaagaaagcagag2880

aagaagggtaaagaagctggtaaaaagaaaggcaaggataacgcagagaaagaggatagt2940

aggtctttgaagaaaaaatcaccttacaaggagaaatctaaagaagaacaagtcaagatc3000

caagaagtaacagaagagtccccccaccaaccagaacctaagataacttacccgtttcac3060

ggatacaatatgggaaatatacccactcaaatctcagggtcaaattactacctgtatcct3120

tctgatggggggcagattgaagtggaaaagacaatgtttgaaaaaggcccaacttttatc3180

aaagtgagagtggtaaaggacaaccacaattttatgattcatttaaatgaccctaaggaa3240

attgtgaaaaaggaagagaaaggggattattatttagaagaggaagaagaaggagatgag3300

gaacaaagtcttgaaacggaagtatcagatgcaaagaataaagctttcagcaagtttgga3360

tctttttctgccaccttagaaaatggaatctgcctctcgataagttactatggatcaaat3420

ggaatggcaccagaagataaggatcctgatttagaaacaatattgaatatcccttcagca3480

ctcactccaacagtggttcctgttatagtgaccgttcctcaaagcaaagctaaagggaaa3540

ataaaaggcaaagaaaaacccaaagaatcccttaaagaagaagaacacccaaaagaagaa3600

gagaaaaaggaagaagaagtagaaccagaacctgttttacaagagactttggatgttccc3660

accttccagagcctaaatgtgtcttgccccagtgggctcctgttgactttcattggacaa3720

gaatctacaggtcaatatgttatagatgaggaacccacctgggacatcatggtccgtcag3780

agctacccccagagggtgaagcactatgagttctataaaacggtgatgccacccgcagag3840

caggaggcttcaagggttatcaccagtcaaggcactgttgtcaaatatatgttggatgga3900

tccacacagattctctttgcagatggtgctgtgagcaggagtcccaattcaggtcttatt3960

tgtcctccttctgaaatgccagcaacgcctcacagtggagatttgatggactctatttct4020

cagcagaaatcagaaacgataccatctgagattaccaacacaaagaaaggaaaaagtcac4080

aaaagtcagtcatcaatggcccataagggtgaaatccatgaccctcctccagaggcagtt4140

caaactgtaactcctgtggaggttcacataggcacctggtttacaaccacacctgaagga4200

aatcggatcggcaccaaaggattagaaagaatagcagacttgaccccattgttatccttt4260

caggccacagatcctgtcaatggaacggttatgacaactcgagaagacaaagttgtcata4320

gttgaaaggaaagatggtactcggatagtggatcatgctgatggtaccagaatcacaacc4380

ttttatcaagtttatgaagatcaaattattctgccagatgatcaagaaacaaccgagggt4440

cctcggactgtcaccaggcaggtgaagtgtatgcgggtagaaagctcacgctatgccact4500

gttatcgccaactgtgaggacagtagctgctgtgccacctttggagatggaacaactatt4560

attgcaaagccacagggaacataccaggtgttacctccaaacacaggctctctttatatt4620

gacaaggattgttcagctgtgtactgccatgagtcaagcagtaatatatactatcctttt4680

caaaagcgtgagcagctgcgagctggcaggtacatcatgaggcatacttcagaggttatc4740

tgtgaggttctggatcctgagggaaacacttttcaggtcatggctgatggtagcatatca4800

actatattacctgaaaaaaaattggaagatgatttaaatgagaaaactgagggctatgat4860

agtctgtcctctatgcaccttgaaaagaatcatcagcaaatctatggtgaacatgtcccc4920

aggttttttgttatgtatgctgatggatcaggaatggaacttcttcgagacagtgacata4980

gaagaatatctatctttggcatataaagaatcaaatactgttgttctccaagagccagtg5040

caggaacagccaggcaccctaaccatcacagtccttcgccctttccatgaagcatcacca5100

tggcaagtaaaaaaggaagatacaattgtccctcctaatctccggtcaaggtcatgggaa5160

acatttccctcagttgagaaaaaaactccaggacctccgtttggtactcagatttggaaa5220

ggcctttgcattgagtccaaacagctagtgagtgccccgggtgccatactcaagagcccc5280

agtgtgctacagatgcgccaattcattcagcatgaggtcataaagaatgaggtgaaactg5340

aggctgcaggtttcccttaaggattacataaactatattctaaagaaagaagatgagctg5400

caggaaatgatggttaaagattccagaactgaggaggagagaggcaatgctgctgatctc5460

ctcaagctggttatgtctttccctaaaatggaggaaactacaaaaagtcatgttactgaa5520

gttgcagctcacctaactgatttattcaagcagtctttggctacgcctccaaaatgccca5580

ccagacacatttggtaaagatttctttgaaaagacatggagacacacagcatcctcaaaa5640

cgctggaaagaaaagatagacaaaacgaggaaggaaattgagacaacacagaattaccta5700

atggatattaagaaccgcataataccacccttttttaaatctgaattgaaccagttatat5760

cagtctcagtataatcacctggacagtctttccaaaaaactgccttcttttacaaagaaa5820

aatgaagatgcaaacgaaacagctgttcaagatacatctgatcttaatctagatttcaag5880

ccacataaggtttcagaacagaaatcctcaagtgtgcctagtcttccaaaaccagagatt5940

tctgcagataagaaggatttcactgctcagaaccaaactgaaaatttaacaaaatctcct6000

gaagaagcagaatcttatgagcccgtgaaaattccaacccagtccttgctgcaggatgtt6060

gcgggacaaacaagaaaagaaaaagtgaagttgcctcattatttgctgagttccaagcct6120

aagtctcaacctcttgcaaaggtgcaagattctgttggaggaaaagtgaacacatcctct6180

gttgcatctgctgccattaataatgcaaagtcatccctttttgggttccatcttctccca6240

tcatcagtcaagtttggagtgcttaaggaaggacatacctatgccacagttgtaaagctc6300

aagaatgttggagtggacttctgcaggtttaaagtaaagcagcccccacccagcacagga6360

ctgaaagtgacttacaaacctggacctgtggcagctggtatgcagacagaactgaatata6420

gagttatttgccacagctgttggagaggatggggccaagggatcagcacacatctctcac6480

aatatcgagattatgacagagcatgaggttctgttcctacctgtggaagcaactgtttta6540

acaagcagcaattatgataaacgaccaaaagactttccccagggaaaagaaaatccaatg6600

gtccagagaacttctacaatttattcctccacacttggagtcttcatgtctcgtaaagtt6660

tctccacattag6672

<210>2

<211>2223

<212>prt

<213>homosapiens

<400>2

metalaprolyslysglulysglyglythrvalasnthrserserlys

151015

iletrpgluproserleuilealaalaglnpheasnglnasnasptrp

202530

glnalaserilealaphevalvalglyasnglnilegluaspaspleu

354045

leuileglnalaleuthrvalalavalglnvalproglnarglysleu

505560

phesermetvalsertrpglnaspileleuglnglnileasngluile

65707580

asnthrleuvalglyseralaserserlyslysalalyslysproval

859095

glyglyasnalaproleutyrtyrgluvalleuthralaalalysala

100105110

ilemetaspserglyglulysleuthrleuproleuileglylysleu

115120125

leulyspheglnleuleuglnilelysphelysaspglnglnargarg

130135140

gluasnglulyslysvalilegluasplysprolysleuglulysasp

145150155160

lysglylysalalysserprolysglulyslysalaproseralalys

165170175

proalalysglylysglylysaspglnproglualaasnalaproval

180185190

lyslysthrthrglnleulysargargglygluaspasphisthrasn

195200205

argtyrileaspaspgluproaspaspglyalaglnhistyrileile

210215220

valvalglypheasnasnproglnleuleualailemetalagluleu

225230235240

glyileproilethrservalilelysilesersergluasntyrglu

245250255

proleuglnthrhisleualaalavalasnglnglnglngluvalleu

260265270

leuglnsergluaspleuglualaglulysleulyslysgluasnala

275280285

ilelysgluleulysthrphetrplystyrleugluprovalleuasn

290295300

asnglulysprogluthrasnleupheaspvalalaargleuglutyr

305310315320

metvallysalaalaasppheproserasptrpseraspglyglumet

325330335

metleulysleuglythraspilephegluasnilealacysleumet

340345350

tyraspileleuasptrplysargglnhisglnhistyrleugluser

355360365

metglnleuileasnvalproglnvalvalasnglulysprovalleu

370375380

glualametprothrserglualaproglnproalavalproalapro

385390395400

glylyslyslysalaglntyrglugluproglnalaproproproval

405410415

thrservalilethrthrgluvalaspmetargtyrtyrasntyrleu

420425430

leuasnproilearggluglupheileservalproleuileleuhis

435440445

cysmetleugluglnvalvalalathrglugluaspleuvalpropro

450455460

serleuarggluproserproargalaaspglyleuasphisargile

465470475480

alaalahisilevalserleuleuproserleucysleusergluarg

485490495

glulyslysasnleuhisaspilepheleusergluglugluasnglu

500505510

serlysalavalprolysglyproleuleuleuasntyrhisaspala

515520525

hisalahislyslystyralaleuglnaspglnlysasnpheasppro

530535540

valglnilegluglnglumetglnserlysleuproleutrpgluphe

545550555560

leuglnpheproleuproproprotrpasnasnthrlysargleuala

565570575

thrilehisgluleumethisphecysthrseraspvalleusertrp

580585590

asngluvalgluargalaphelysvalphethrphegluserleulys

595600605

leusergluvalaspglulysglylysleulysproserglymetmet

610615620

cysglyseraspserglumetpheasnileprotrpaspasnproala

625630635640

argphealalysglnileargglnglntyrvalmetlysmetasnthr

645650655

glnglualalysglnlysalaaspilelysilelysaspargthrleu

660665670

phevalaspglnasnleusermetservalglnaspasngluserasn

675680685

arggluproseraspproserglncysaspalaasnasnmetlyshis

690695700

seraspleuasnasnleulysleuservalproaspasnargglnleu

705710715720

leugluglngluserilemetlysalaglnproglnhisgluserleu

725730735

gluglnthrthrasnasngluilelysaspaspalavalthrlysala

740745750

aspserhisglulyslysprolyslysmetmetvalglualaaspleu

755760765

gluaspilelyslysthrglnglnargserleumetasptrpserphe

770775780

thrgluhisphelysprolysvalleuleuglnvalleuglngluala

785790795800

hislysglntyrargcysvalaspsertyrtyrhisthrglnaspasn

805810815

serleuleuleuvalphehisasnprometasnargglnargleuhis

820825830

cysglutyrtrpasnilealaleuhisserasnvalglypheargasn

835840845

tyrleugluleuvalalalysserileglnasptrpilethrlysglu

850855860

glualailetyrglngluserlysmetasnglulysileileargthr

865870875880

argalagluleugluleulysserseralaasnalalysleuthrser

885890895

alaserlysilepheserilelysgluserlysserasnlysglyile

900905910

serlysthrgluileseraspglnglulysglulysglulysglulys

915920925

ilepropheileleugluglyserleulysalatrplysgluglugln

930935940

hisargleualagluglugluargleuarggluglulyslysalaglu

945950955960

lyslysglylysglualaglylyslyslysglylysaspasnalaglu

965970975

lysgluaspserargserleulyslyslysserprotyrlysglulys

980985990

serlysglugluglnvallysileglngluvalthrglugluserpro

99510001005

hisglnprogluprolysilethrtyrprophehisglytyrasn

101010151020

metglyasnileprothrglnileserglyserasntyrtyrleu

102510301035

tyrproseraspglyglyglnilegluvalglulysthrmetphe

104010451050

glulysglyprothrpheilelysvalargvalvallysaspasn

105510601065

hisasnphemetilehisleuasnaspprolysgluilevallys

107010751080

lysgluglulysglyasptyrtyrleuglugluglugluglugly

108510901095

aspglugluglnserleugluthrgluvalseraspalalysasn

110011051110

lysalapheserlyspheglyserpheseralathrleugluasn

111511201125

glyilecysleuserilesertyrtyrglyserasnglymetala

113011351140

progluasplysaspproaspleugluthrileleuasnilepro

114511501155

seralaleuthrprothrvalvalprovalilevalthrvalpro

116011651170

glnserlysalalysglylysilelysglylysglulysprolys

117511801185

gluserleulysgluglugluhisprolysglugluglulyslys

119011951200

gluglugluvalgluprogluprovalleuglngluthrleuasp

120512101215

valprothrpheglnserleuasnvalsercysproserglyleu

122012251230

leuleuthrpheileglyglngluserthrglyglntyrvalile

123512401245

aspglugluprothrtrpaspilemetvalargglnsertyrpro

125012551260

glnargvallyshistyrgluphetyrlysthrvalmetpropro

126512701275

alagluglnglualaserargvalilethrserglnglythrval

128012851290

vallystyrmetleuaspglyserthrglnileleuphealaasp

129513001305

glyalavalserargserproasnserglyleuilecyspropro

131013151320

serglumetproalathrprohisserglyaspleumetaspser

132513301335

ileserglnglnlyssergluthrileprosergluilethrasn

134013451350

thrlyslysglylysserhislysserglnsersermetalahis

135513601365

lysglygluilehisaspproproproglualavalglnthrval

137013751380

thrprovalgluvalhisileglythrtrpphethrthrthrpro

138513901395

gluglyasnargileglythrlysglyleugluargilealaasp

140014051410

leuthrproleuleuserpheglnalathraspprovalasngly

141514201425

thrvalmetthrthrarggluasplysvalvalilevalgluarg

143014351440

lysaspglythrargilevalasphisalaaspglythrargile

144514501455

thrthrphetyrglnvaltyrgluaspglnileileleuproasp

146014651470

aspglngluthrthrgluglyproargthrvalthrargglnval

147514801485

lyscysmetargvalgluserserargtyralathrvalileala

149014951500

asncysgluaspsersercyscysalathrpheglyaspglythr

150515101515

thrileilealalysproglnglythrtyrglnvalleupropro

152015251530

asnthrglyserleutyrileasplysaspcysseralavaltyr

153515401545

cyshisgluserserserasniletyrtyrpropheglnlysarg

155015551560

gluglnleuargalaglyargtyrilemetarghisthrserglu

156515701575

valilecysgluvalleuaspprogluglyasnthrpheglnval

158015851590

metalaaspglyserileserthrileleuproglulyslysleu

159516001605

gluaspaspleuasnglulysthrgluglytyraspserleuser

161016151620

sermethisleuglulysasnhisglnglniletyrglygluhis

162516301635

valproargphephevalmettyralaaspglyserglymetglu

164016451650

leuleuargaspseraspilegluglutyrleuserleualatyr

165516601665

lysgluserasnthrvalvalleuglngluprovalglnglugln

167016751680

proglythrleuthrilethrvalleuargprophehisgluala

168516901695

serprotrpglnvallyslysgluaspthrilevalproproasn

170017051710

leuargserargsertrpgluthrpheproservalglulyslys

171517201725

thrproglypropropheglythrglniletrplysglyleucys

173017351740

ilegluserlysglnleuvalseralaproglyalaileleulys

174517501755

serproservalleuglnmetargglnpheileglnhisgluval

176017651770

ilelysasngluvallysleuargleuglnvalserleulysasp

177517801785

tyrileasntyrileleulyslysgluaspgluleuglnglumet

179017951800

metvallysaspserargthrgluglugluargglyasnalaala

180518101815

aspleuleulysleuvalmetserpheprolysmetglugluthr

182018251830

thrlysserhisvalthrgluvalalaalahisleuthraspleu

183518401845

phelysglnserleualathrproprolyscysproproaspthr

185018551860

pheglylysaspphepheglulysthrtrparghisthralaser

186518701875

serlysargtrplysglulysileasplysthrarglysgluile

188018851890

gluthrthrglnasntyrleumetaspilelysasnargileile

189519001905

proprophephelyssergluleuasnglnleutyrglnsergln

191019151920

tyrasnhisleuaspserleuserlyslysleuproserphethr

192519301935

lyslysasngluaspalaasngluthralavalglnaspthrser

194019451950

aspleuasnleuaspphelysprohislysvalsergluglnlys

195519601965

serserservalproserleuprolysprogluileseralaasp

197019751980

lyslysaspphethralaglnasnglnthrgluasnleuthrlys

198519901995

serprogluglualaglusertyrgluprovallysileprothr

200020052010

glnserleuleuglnaspvalalaglyglnthrarglysglulys

201520202025

vallysleuprohistyrleuleuserserlysprolyssergln

203020352040

proleualalysvalglnaspservalglyglylysvalasnthr

204520502055

serservalalaseralaalaileasnasnalalysserserleu

206020652070

pheglyphehisleuleuproserservallyspheglyvalleu

207520802085

lysgluglyhisthrtyralathrvalvallysleulysasnval

209020952100

glyvalaspphecysargphelysvallysglnproproproser

210521102115

thrglyleulysvalthrtyrlysproglyprovalalaalagly

212021252130

metglnthrgluleuasnilegluleuphealathralavalgly

213521402145

gluaspglyalalysglyseralahisileserhisasnileglu

215021552160

ilemetthrgluhisgluvalleupheleuprovalglualathr

216521702175

valleuthrserserasntyrasplysargprolysaspphepro

218021852190

glnglylysgluasnprometvalglnargthrserthriletyr

219522002205

serserthrleuglyvalphemetserarglysvalserprohis

221022152220

<210>3

<211>20

<212>dna

<213>人工序列

<400>3

tggtgccatcttcgtattca20

<210>4

<211>20

<212>dna

<213>人工序列

<400>4

atagaccctgctttggatgg20