本发明涉及致病基因，尤其是涉及一种严重弱精子症新致病基因及其应用。
背景技术：
严重弱精子症是一种较常见的导致男性不育的畸形精子症，是一种与遗传、生活方式等因素有关的疾病，其主要临床特征是精子运动能力的严重丧失。国外早期的分子方面发病机制研究发现akap3、akap4、slc26a8、dnah1、catsper2、galntl5等基因发生了突变(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.)，并导致精子尾部形态及运动异常。
目前严重弱精子症的基因突变谱尚未完全发现，基因型和表现型的关系也不明确。目前单基因疾病的研究开始大量采用全外显子组测序(whole-exomesequencing)及全基因组测序(whole-genomesequencing)的方法，这两种方法被成功的应用于发现稀有单基因疾病的致病基因。全外显子组测序及全基因组测序技术已被证明为降低稀有单基因疾病候选基因甚至发现其致病基因的有力、有效手段。仅通过对几个很少的个体(包括患者及正常对照)的全外显子组或基因组进行测序来筛选与疾病相关的变异，其成功率大为提升。
因此本领域对严重弱精子症的研究尚不清晰，对造成该疾病的原因更不明了，因此本领域迫切需要对严重弱精子症的致病机理进行研究，找到新致病基因及突变位点。
技术实现要素：
本发明的第一目的在于提供通过外显子组测序的方法确定严重弱精子症的新致病基因。
本发明的第二目的在于提供严重弱精子症的生物标记物。
本发明的第三目的在于提供突变的spag17基因。
本发明的第四目的在于提供突变的spag17蛋白。
本发明的第五目的在于提供一种检测严重弱精子症的方法。
本发明的第六目的在于提供通过pcr检测spag17基因或spag17蛋白突变中使用的引物对。
本发明的第七目的在于提供与突变spag17基因互补的核酸探针。
本发明的第八目的在于提供检测突变spag17基因或spag17蛋白的试剂盒。
本发明的第九目的在于提供检测突变spag17基因的试剂盒。
本发明的第十目的在于提供一种严重弱精子症新致病基因的应用。
所述通过外显子组测序的方法确定严重弱精子症的新致病基因。
所述严重弱精子症的生物标记物，即突变的spag17基因或spag17蛋白，所述生物标记物是具有选自如下的突变的spag17基因或spag17蛋白：
在外显子30中错义突变(c.4343g>a；p.r1448q)。
所述突变的spag17基因为seqidno:1的序列中具有以下突变：
在外显子30中错义突变(c.4343g>a)。
所述突变的spag17蛋白为seqidno:2的序列中具有以下突变：
在外显子30中错义突变(p.r1448q)。
所述一种检测严重弱精子症的方法，包括检测受试者的spag17基因或spag17蛋白中是否存在突变位点，若有突变位点，则所述受试者被鉴定为患有严重弱精子症或易患严重弱精子症，所述突变位点选自如下一种：
在外显子30中错义突变(c.4343g>a；p.r1448q)。
所述spag17蛋白为seqidno:2的序列表示。
所述spag17基因为seqidno:1的序列表示。
所述检测严重弱精子症的方法包括如下至少一组引物扩增的步骤：
seqidno:3和seqidno:4。
所述检测严重弱精子症的方法中检测突变位点通过选自如下进行：测序、电泳、核酸杂交、原位杂交、pcr、逆转录酶链反应和变性高效液相色谱。
所述通过pcr检测spag17基因或spag17蛋白突变中使用的引物对，所述突变是选自如下一种：
在外显子30中错义突变(c.4343g>a；p.r1448q)，
其中所述引物对分别基于选自如下的位置前后设计，使得扩增该位置(编号基于spag17的cdna序列)：4343。
所述与突变spag17基因互补的核酸探针，所述突变是选自如下一种：
在外显子30中错义突变(c.4343g>a；p.r1448q)，
所述探针与突变spag17基因的互补区包括选自如下的位置(编号基于spag17的cdna序列)：4343。
所述检测突变spag17基因或spag17蛋白的试剂盒，包含一组或多组引物对，其中所述突变是选自如下一种：
在外显子30中错义突变(c.4343g>a；p.r1448q)，
其中所述引物对分别基于选自如下的位置前后设计，使得其扩增产物涵盖该位置(编号基于spag17的cdna序列)：4343。
所述检测突变spag17基因或spag17蛋白的试剂盒包含选自如下的至少一组引物：
seqidno:3和seqidno:4。
所述检测突变spag17基因的试剂盒，包含一个或多个核酸探针，所述突变是选自如下一种：
在外显子30中错义突变(c.4343g>a；p.r1448q)，
所述探针与突变spag17基因上包含选自如下的位置的区域互补(编号基于spag17的cdna序列)：4343。
所述一种严重弱精子症新致病基因的应用。
本发明将为严重弱精子症的发病机制研究奠定重要基础，有可能为严重弱精子症的患者治疗提供全新的理论依据。
具体实施方式
在本发明中，采用本领域通用表示法表示突变。例如，突变(c.4343g>a；p.r1448q)中，c表示cdna，p表示蛋白质，dna水平的突变对应蛋白质水平的突变。
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，本领域技术人员应当理解，实际包括互补双链的任意一条，或者两条。虽然多数情况下至给出了一条链，但实际上也公开了与之互补的另一条链。例如提及spag17基因的cdna序列，实际上包括该序列以及其互补序列。例如，提及seqidno:1，实际包括其互补序列。本领域技术人员还可以理解，利用一条链可以检测另一条链，反之亦然。
本申请中的基因序列包括dna形式或rna形式，公开其中一种，意味着另一种也被公开。例如提及spag17基因的cdna序列，实际也包括相应的rna序列。
实施例1确定严重弱精子症的致病基因。
本发明收集1个近亲婚配家系中的两位双胞胎患有严重弱精子症的病例，两位患者均表现为不育。先证者29岁，父母为三代内姑表亲结婚，生育3男1女，哥哥生育1子，妹妹生育两胎，双胞胎弟弟不育。先证者外生殖器发育正常，双侧睾丸大小正常，双侧精索静脉未触及异常。外周血染色体核型分析正常，y染色体微缺失检测未发现异常。无支气管扩张等上呼吸道疾病，无内脏转位，无鼻窦炎病史。在我院三次精液检查平均值为精液量2.8ml，精子密度27.5百万/ml，前向运动精子占比1.8％，非前向运动精子1.6％。精子形态改良巴氏染色读取显示正常形态精子4.5％，精子顶体酶28miu/ml，精浆生化果糖、中性糖苷酶活性及精浆锌均未见异常，根据以上检查结果提示患者为严重弱精子症。
本发明对先症者进行了全外显子组测序，具体步骤如下：
样品制备：采集所述患者及其父母家人外周血，利用试剂盒抽提外周血白细胞中的基因组dna(qiaampdnaminikit51304.qiagen,usa)，利用nanodrop2000测量dna的浓度及纯度(thermoscientific,usa)，所得的每个标本基因组dna的od260/od280均位于1.7～2.0之间，浓度不少于100ng/μl，总量不少于30μl。
然后，对上述1个样本的外显子组序列进行了测序。测序平台为illuminahiseq2000，依照illumina标准建库说明书(参见http://www.illumina.com/)进行测序，简述如下：
1)使用illumina公司试剂盒truseqdnalibraryprepkit制作dna文库。在外显子捕获过程中使用agilent公司试剂盒sureselecthumanallexonv5；
2)使用illumina公司hiseq2000测序平台对捕获的外显子区域进行平行测序读取长度为90bp，每个样本的平均测序深度最少为100；
3)使用burrows-wheeleraligner(http://bio-bwa.sourceforge.net/)将测序结果比对到人参考基因组中(hg19)。使用samtools将重复读取结果去除，通过gatk(https://www.broadinstitute.org/gatk/)进行调整和校准。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。
将等位基因频率大于1％的突变位点剔除(参考数据库包括dbsnp，1000genomes，exomeaggregationconsortium(exac))。
发现先症者在基因spag17上具有纯合突变：通过家系中2名患者和4位正常人snp筛查共分离，这些位点可能是致病位点。
结果显示于表3。
因此本发明认为spag17基因极有可能为严重弱精子症的致病基因。
实施例2在其他病例中确认上述严重弱精子症的致病基因。
本发明纳入所有严重弱精子症患者，对spag17基因为严重弱精子症的致病基因进行验证。验证中利用sanger法测序验证spag17基因的突变，其中考虑了家系内等样本验证情况。
分别对2名患者、4名家系内正常人(即其中2名患者的父母及哥哥姐姐，他们均未发病)基因进行检测。
具体方法步骤如下：
1.dna提取：
分别提取2名患者、4名家系内正常人的外周静脉血，按照实施例1的方法提取基因组dna和测定dna含量。
2.引物设计及pcr反应
引物设计参考人类基因组序列数据库hg19，引物序列参见表1。
表1
pcr扩增体系参见表2。
表2
pcr反应条件：
94℃变性5min，然后进入每个循环，即94℃变性30s，退火(通常为55℃，根据不同引物的退火温度设定)30s，72℃延伸1min，总共设定30个循环。所有循环结束，最终72℃延伸10min，并将pcr产物保存在4或-20℃。
3)将步骤2)中获得的获自2名患者、4名家系内正常人(即该2例患者的父母，他们均未发病)的pcr扩征产物直接进行dna测序，方法同实施例1。
本发明收集到的家系内2例严重弱精子症病例均在spag17基因中检测到有意义的突变位点。同时所发现的突变位点在家族正常人中均未能检测到。这些突变位点在exac数据库中的频率极低或无频率(见表3)，提示所检测到的位点很可能并非snp。因此认为spag17基因为严重弱精子症的致病基因。
表3
序列表说明
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>厦门市妇幼保健院（厦门市计划生育服务中心），首都医科大学附属北京妇产医院，烟台毓璜顶医院
<120>一种严重弱精子症新致病基因及其应用
<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