CN114085900B

CN114085900B - 与遗传性肾小球疾病相关的突变基因及其应用

Info

Publication number: CN114085900B
Application number: CN202111382255.5A
Authority: CN
Inventors: 刘哲; 梁庆渊; 赵娜娜; 赖开生; 刘昕超; 高璇; 李方玉; 侯青; 惠汝太
Original assignee: Bosinor Beijing Medical Laboratory Co ltd
Current assignee: Bosinor Beijing Medical Laboratory Co ltd
Priority date: 2021-11-22
Filing date: 2021-11-22
Publication date: 2022-07-29
Anticipated expiration: 2041-11-22
Also published as: CN114085900A

Abstract

本发明涉及人类遗传学和内科心血管技术领域，具体涉及了与遗传性肾小球疾病相关的突变基因，与野生型FN1基因编码DNA的参考序列相比，突变基因的核苷酸序列为SEQ ID NO:3；在基因组位置chr2:216273077处，碱基A突变为碱基G；参考基因组版本是GRCh37。本发明还涉及上述与遗传性肾小球疾病相关的突变基因在制备检测试剂盒中的应用。本发明提供的突变基因可以作为临床辅助诊断的生物标志物；检测该变异的携带者，为受试者提供优生优育指导和遗传咨询，减少患儿出生，对纤维连接蛋白肾小球病的早期诊断，或者辅助临床判断具有重要意义。

Description

与遗传性肾小球疾病相关的突变基因及其应用

技术领域

本发明涉及人类遗传学和内科心血管技术领域，尤其涉及与遗传性肾小球疾病相关的突变基因及其应用。

背景技术

纤维连接蛋白肾小球病(fibronectin glomerulopathy,FNG)于1987年由Tuttle等首先报道是一种具有家族遗传性的常染色体显性遗传性肾小球疾病，主要临床表现为：肾性高血压，蛋白尿，镜下血尿，肾病综合征，肾功能衰竭，终末期肾病，肾小球扩张，系膜和内皮下颗粒状或纤维性沉积物，发病年龄3～65岁，年轻人多见，男女均可受累，文献报道84％的患者首发症状在50岁之前，老年发病者多为女性。目前尚无特异性治疗，一般多给予对症治疗。Castelletti等报告40％FNG患者携带突变基因FN1。

FN1基因编码纤维连接蛋白，在细胞表面和细胞外基质中以二聚体或多聚体形式存在。纤维连接蛋白参与细胞粘附和迁移过程，包括胚胎形成，伤口愈合，凝血，宿主防御和转移。

目前已发现大量FN1基因突变位点，例如c.3452C>T、c.2674T>A、c.2638T>C等等，但是仍存在未知的FN1基因突变位点，进一步发现新的FN1基因的突变位点对于研究纤维连接蛋白肾小球病的发病机制，对纤维连接蛋白肾小球病的早期诊断，或者辅助临床判断具有重要意义。

发明内容

本发明的目的是针对纤维连接蛋白肾小球病，提供一种与遗传性肾小球疾病相关的突变基因及其应用。

本发明提供的技术方案如下：

一、本发明提供与遗传性肾小球疾病相关的突变基因，与野生型FN1基因编码DNA的参考序列相比，突变基因的核苷酸序列为SEQ ID NO:3；在基因组位置chr2:216273077处，碱基A突变为碱基G；参考基因组版本是GRCh37。

二、本发明还提供了上述与遗传性肾小球疾病相关的突变基因在制备检测试剂盒中的应用。

优选地，所述纤维连接蛋白肾小球病检测试剂盒包括引物SEQ ID NO:1和SEQ IDNO:2。

优选地，所述纤维连接蛋白肾小球病检测试剂盒还包括Taq DNA聚合酶和PCR缓冲液。

三、本发明的原理和有益效果在于：

本发明公开的突变基因可以作为临床辅助诊断的生物标志物，对纤维连接蛋白肾小球病的早期诊断，或者辅助临床判断具有重要意义；基于与遗传性肾小球疾病相关的突变基因开发的检测试剂盒，可以检测出具有纤维连接蛋白肾小球病突变基因的患者，为受试者提供优生优育指导和遗传咨询，减少患儿出生。

附图说明

图1为实施例3的家系图；

图2为实施例3中先证者、先证者母亲等人的Sanger测序图；

图3为实施例3家系中先证者父亲、先证者妹妹等人的Sanger测序图。

具体实施方式

下面通过具体实施方式进一步详细说明：

实施例1-与遗传性肾小球疾病相关的突变基因

与遗传性肾小球疾病相关的突变基因，具体突变如下表1所示：

表1与遗传性肾小球疾病相关的突变基因的具体检测结果

基因

基因组位置

转录本号

碱基改变

氨基酸改变

参考基因组版本

外显子号

FN1

chr2:216273077

NM_001306129

c.2372A>G

p.Tyr791Cys

GRCh37/hg19

Exon16

(1)在基因组位置chr2:216273076-chr2:216273125处，野生型FN1基因的序列为：

TGAACATCCCTGACCTGCTTCCTGGCCGAAAATACATTGTAAATGTCT

T，

为野生型FN1基因在基因组chr2:216273077处的碱基。

在对应基因组位置处，与遗传性肾小球疾病相关的突变基因的序列为：

TGAACATCCCTGACCTGCTTCCTGGCCGAAAATACATTGTAAATGTCT

T，

为突变基因FN1在基因组chr2:216273077处的碱基。

(2)转录本号为NM_001306129时，野生型FN1基因编码DNA的参考序列为：

ATGCTTAGGGGTCCGGGGCCCGGGCTGCTGCTGCTGGCCGTCCAGTGCCTGGGGACAGCGGTGCCCTCCACGGGAGCCTCGAAGAGCAAGAGGCAGGCTCAGCAAATGGTTCAGCCCCAGTCCCCGGTGGCTGTCAGTCAAAGCAAGCCCGGTTGTTATGACAATGGAAAACACTATCAGATAAATCAACAGTGGGAGCGGACCTACCTAGGCAATGCGTTGGTTTGTACTTGTTATGGAGGAAGCCGAGGTTTTAACTGCGAGAGTAAACCTGAAGCTGAAGAGACTTGCTTTGACAAGTACACTGGGAACACTTACCGAGTGGGTGACACTTATGAGCGTCCTAAAGACTCCATGATCTGGGACTGTACCTGCATCGGGGCTGGGCGAGGGAGAATAAGCTGTACCATCGCAAACCGCTGCCATGAAGGGGGTCAGTCCTACAAGATTGGTGACACCTGGAGGAGACCACATGAGACTGGTGGTTACATGTTAGAGTGTGTGTGTCTTGGTAATGGAAAAGGAGAATGGACCTGCAAGCCCATAGCTGAGAAGTGTTTTGATCATGCTGCTGGGACTTCCTATGTGGTCGGAGAAACGTGGGAGAAGCCCTACCAAGGCTGGATGATGGTAGATTGTACTTGCCTGGGAGAAGGCAGCGGACGCATCACTTGCACTTCTAGAAATAGATGCAACGATCAGGACACAAGGACATCCTATAGAATTGGAGACACCTGGAGCAAGAAGGATAATCGAGGAAACCTGCTCCAGTGCATCTGCACAGGCAACGGCCGAGGAGAGTGGAAGTGTGAGAGGCACACCTCTGTGCAGACCACATCGAGCGGATCTGGCCCCTTCACCGATGTTCGTGCAGCTGTTTACCAACCGCAGCCTCACCCCCAGCCTCCTCCCTATGGCCACTGTGTCACAGACAGTGGTGTGGTCTACTCTGTGGGGATGCAGTGGCTGAAGACACAAGGAAATAAGCAAATGCTTTGCACGTGCCTGGGCAACGGAGTCAGCTGCCAAGAGACAGCTGTAACCCAGACTTACGGTGGCAACTCAAATGGAGAGCCATGTGTCTTACCATTCACCTACAATGGCAGGACGTTCTACTCCTGCACCACAGAAGGGCGACAGGACGGACATCTTTGGTGCAGCACAACTTCGAATTATGAGCAGGACCAGAAATACTCTTTCTGCACAGACCACACTGTTTTGGTTCAGACTCGAGGAGGAAATTCCAATGGTGCCTTGTGCCACTTCCCCTTCCTATACAACAACCACAATTACACTGATTGCACTTCTGAGGGCAGAAGAGACAACATGAAGTGGTGTGGGACCACACAGAACTATGATGCCGACCAGAAGTTTGGGTTCTGCCCCATGGCTGCCCACGAGGAAATCTGCACAACCAATGAAGGGGTCATGTACCGCATTGGAGATCAGTGGGATAAGCAGCATGACATGGGTCACATGATGAGGTGCACGTGTGTTGGGAATGGTCGTGGGGAATGGACATGCATTGCCTACTCGCAGCTTCGAGATCAGTGCATTGTTGATGACATCACTTACAATGTGAACGACACATTCCACAAGCGTCATGAAGAGGGGCACATGCTGAACTGTACATGCTTCGGTCAGGGTCGGGGCAGGTGGAAGTGTGATCCCGTCGACCAATGCCAGGATTCAGAGACTGGGACGTTTTATCAAATTGGAGATTCATGGGAGAAGTATGTGCATGGTGTCAGATACCAGTGCTACTGCTATGGCCGTGGCATTGGGGAGTGGCATTGCCAACCTTTACAGACCTATCCAAGCTCAAGTGGTCCTGTCGAAGTATTTATCACTGAGACTCCGAGTCAGCCCAACTCCCACCCCATCCAGTGGAATGCACCACAGCCATCTCACATTTCCAAGTACATTCTCAGGTGGAGACCTAAAAATTCTGTAGGCCGTTGGAAGGAAGCTACCATACCAGGCCACTTAAACTCCTACACCATCAAAGGCCTGAAGCCTGGTGTGGTATACGAGGGCCAGCTCATCAGCATCCAGCAGTACGGCCACCAAGAAGTGACTCGCTTTGACTTCACCACCACCAGCACCAGCACACCTGTGACCAGCAACACCGTGACAGGAGAGACGACTCCCTTTTCTCCTCTTGTGGCCACTTCTGAATCTGTGACCGAAATCACAGCCAGTAGCTTTGTGGTCTCCTGGGTCTCAGCTTCCGACACCGTGTCGGGATTCCGGGTGGAATATGAGCTGAGTGAGGAGGGAGATGAGCCACAGTACCTGGATCTTCCAAGCACAGCCACTTCTGTGAACATCCCTGACCTGCTTCCTGGCCGAAAATACATTGTAAATGTCT

TCAGATATCTGAGGATGGGGAGCAGAGTTTGATCCTGTCTACTTCACAAACAACAGCGCCTGATGCCCCTCCTGACACGACTGTGGACCAAGTTGATGACACCTCAATTGTTGTTCGCTGGAGCAGACCCCAGGCTCCCATCACAGGGTACAGAATAGTCTATTCGCCATCAGTAGAAGGTAGCAGCACAGAACTCAACCTTCCTGAAACTGCAAACTCCGTCACCCTCAGTGACTTGCAACCTGGTGTTCAGTATAACATCACTATCTATGCTGTGGAAGAAAATCAAGAAAGTACACCTGTTGTCATTCAACAAGAAACCACTGGCACCCCACGCTCAGATACAGTGCCCTCTCCCAGGGACCTGCAGTTTGTGGAAGTGACAGACGTGAAGGTCACCATCATGTGGACACCGCCTGAGAGTGCAGTGACCGGCTACCGTGTGGATGTGATCCCCGTCAACCTGCCTGGCGAGCACGGGCAGAGGCTGCCCATCAGCAGGAACACCTTTGCAGAAGTCACCGGGCTGTCCCCTGGGGTCACCTATTACTTCAAAGTCTTTGCAGTGAGCCATGGGAGGGAGAGCAAGCCTCTGACTGCTCAACAGACAACCAAACTGGATGCTCCCACTAACCTCCAGTTTGTCAATGAAACTGATTCTACTGTCCTGGTGAGATGGACTCCACCTCGGGCCCAGATAACAGGATACCGACTGACCGTGGGCCTTACCCGAAGAGGACAGCCCAGGCAGTACAATGTGGGTCCCTCTGTCTCCAAGTACCCACTGAGGAATCTGCAGCCTGCATCTGAGTACACCGTATCCCTCGTGGCCATAAAGGGCAACCAAGAGAGCCCCAAAGCCACTGGAGTCTTTACCACACTGCAGCCTGGGAGCTCTATTCCACCTTACAACACCGAGGTGACTGAGACCACCATTGTGATCACATGGACGCCTGCTCCAAGAATTGGTTTTAAGCTGGGTGTACGACCAAGCCAGGGAGGAGAGGCACCACGAGAAGTGACTTCAGACTCAGGAAGCATCGTTGTGTCCGGCTTGACTCCAGGAGTAGAATACGTCTACACCATCCAAGTCCTGAGAGATGGACAGGAAAGAGATGCGCCAATTGTAAACAAAGTGGTGACACCATTGTCTCCACCAACAAACTTGCATCTGGAGGCAAACCCTGACACTGGAGTGCTCACAGTCTCCTGGGAGAGGAGCACCACCCCAGACATTACTGGTTATAGAATTACCACAACCCCTACAAACGGCCAGCAGGGAAATTCTTTGGAAGAAGTGGTCCATGCTGATCAGAGCTCCTGCACTTTTGATAACCTGAGTCCCGGCCTGGAGTACAATGTCAGTGTTTACACTGTCAAGGATGACAAGGAAAGTGTCCCTATCTCTGATACCATCATCCCAGAGGTGCCCCAACTCACTGACCTAAGCTTTGTTGATATAACCGATTCAAGCATCGGCCTGAGGTGGACCCCGCTAAACTCTTCCACCATTATTGGGTACCGCATCACAGTAGTTGCGGCAGGAGAAGGTATCCCTATTTTTGAAGATTTTGTGGACTCCTCAGTAGGATACTACACAGTCACAGGGCTGGAGCCGGGCATTGACTATGATATCAGCGTTATCACTCTCATTAATGGCGGCGAGAGTGCCCCTACTACACTGACACAACAAACGGCTGTTCCTCCTCCCACTGACCTGCGATTCACCAACATTGGTCCAGACACCATGCGTGTCACCTGGGCTCCACCCCCATCCATTGATTTAACCAACTTCCTGGTGCGTTACTCACCTGTGAAAAATGAGGAAGATGTTGCAGAGTTGTCAATTTCTCCTTCAGACAATGCAGTGGTCTTAACAAATCTCCTGCCTGGTACAGAATATGTAGTGAGTGTCTCCAGTGTCTACGAACAACATGAGAGCACACCTCTTAGAGGAAGACAGAAAACAGGTCTTGATTCCCCAACTGGCATTGACTTTTCTGATATTACTGCCAACTCTTTTACTGTGCACTGGATTGCTCCTCGAGCCACCATCACTGGCTACAGGATCCGCCATCATCCCGAGCACTTCAGTGGGAGACCTCGAGAAGATCGGGTGCCCCACTCTCGGAATTCCATCACCCTCACCAACCTCACTCCAGGCACAGAGTATGTGGTCAGCATCGTTGCTCTTAATGGCAGAGAGGAAAGTCCCTTATTGATTGGCCAACAATCAACAGTTTCTGATGTTCCGAGGGACCTGGAAGTTGTTGCTGCGACCCCCACCAGCCTACTGATCAGCTGGGATGCTCCTGCTGTCACAGTGAGATATTACAGGATCACTTACGGAGAGACAGGAGGAAATAGCCCTGTCCAGGAGTTCACTGTGCCTGGGAGCAAGTCTACAGCTACCATCAGCGGCCTTAAACCTGGAGTTGATTATACCATCACTGTGTATGCTGTCACTGGCCGTGGAGACAGCCCCGCAAGCAGCAAGCCAATTTCCATTAATTACCGAACAGAAATTGACAAACCATCCCAGATGCAAGTGACCGATGTTCAGGACAACAGCATTAGTGTCAAGTGGCTGCCTTCAAGTTCCCCTGTTACTGGTTACAGAGTAACCACCACTCCCAAAAATGGACCAGGACCAACAAAAACTAAAACTGCAGGTCCAGATCAAACAGAAATGACTATTGAAGGCTTGCAGCCCACAGTGGAGTATGTGGTTAGTGTCTATGCTCAGAATCCAAGCGGAGAGAGTCAGCCTCTGGTTCAGACTGCAGTAACCAACATTGATCGCCCTAAAGGACTGGCATTCACTGATGTGGATGTCGATTCCATCAAAATTGCTTGGGAAAGCCCACAGGGGCAAGTTTCCAGGTACAGGGTGACCTACTCGAGCCCTGAGGATGGAATCCATGAGCTATTCCCTGCACCTGATGGTGAAGAAGACACTGCAGAGCTGCAAGGCCTCAGACCGGGTTCTGAGTACACAGTCAGTGTGGTTGCCTTGCACGATGATATGGAGAGCCAGCCCCTGATTGGAACCCAGTCCACAGCTATTCCTGCACCAACTGACCTGAAGTTCACTCAGGTCACACCCACAAGCCTGAGCGCCCAGTGGACACCACCCAATGTTCAGCTCACTGGATATCGAGTGCGGGTGACCCCCAAGGAGAAGACCGGACCAATGAAAGAAATCAACCTTGCTCCTGACAGCTCATCCGTGGTTGTATCAGGACTTATGGTGGCCACCAAATATGAAGTGAGTGTCTATGCTCTTAAGGACACTTTGACAAGCAGACCAGCTCAGGGAGTTGTCACCACTCTGGAGAATGTCAGCCCACCAAGAAGGGCTCGTGTGACAGATGCTACTGAGACCACCATCACCATTAGCTGGAGAACCAAGACTGAGACGATCACTGGCTTCCAAGTTGATGCCGTTCCAGCCAATGGCCAGACTCCAATCCAGAGAACCATCAAGCCAGATGTCAGAAGCTACACCATCACAGGTTTACAACCAGGCACTGACTACAAGATCTACCTGTACACCTTGAATGACAATGCTCGGAGCTCCCCTGTGGTCATCGACGCCTCCACTGCCATTGATGCACCATCCAACCTGCGTTTCCTGGCCACCACACCCAATTCCTTGCTGGTATCATGGCAGCCGCCACGTGCCAGGATTACCGGCTACATCATCAAGTATGAGAAGCCTGGGTCTCCTCCCAGAGAAGTGGTCCCTCGGCCCCGCCCTGGTGTCACAGAGGCTACTATTACTGGCCTGGAACCGGGAACCGAATATACAATTTATGTCATTGCCCTGAAGAATAATCAGAAGAGCGAGCCCCTGATTGGAAGGAAAAAGACAGACGAGCTTCCCCAACTGGTAACCCTTCCACACCCCAATCTTCATGGACCAGAGATCTTGGATGTTCCTTCCACAGTTCAAAAGACCCCTTTCGTCACCCACCCTGGGTATGACACTGGAAATGGTATTCAGCTTCCTGGCACTTCTGGTCAGCAACCCAGTGTTGGGCAACAAATGATCTTTGAGGAACATGGTTTTAGGCGGACCACACCGCCCACAACGGCCACCCCCATAAGGCATAGGCCAAGACCATACCCGCCGAATGTAGGACAAGAAGCTCTCTCTCAGACAACCATCTCATGGGCCCCATTCCAGGACACTTCTGAGTACATCATTTCATGTCATCCTGTTGGCACTGATGAAGAACCCTTACAGTTCAGGGTTCCTGGAACTTCTACCAGTGCCACTCTGACAGGCCTCACCAGAGGTGCCACCTACAACGTCATAGTGGAGGCACTGAAAGACCAGCAGAGGCATAAGGTTCGGGAAGAGGTTGTTACCGTGGGCAACTCTGTCAACGAAGGCTTGAACCAACCTACGGATGACTCGTGCTTTGACCCCTACACAGTTTCCCATTATGCCGTTGGAGATGAGTGGGAACGAATGTCTGAATCAGGCTTTAAACTGTTGTGCCAGTGCTTAGGCTTTGGAAGTGGTCATTTCAGATGTGATTCATCTAGATGGTGCCATGACAATGGTGTGAACTACAAGATTGGAGAGAAGTGGGACCGTCAGGGAGAAAATGGCCAGATGATGAGCTGCACATGTCTTGGGAACGGAAAAGGAGAATTCAAGTGTGACCCTCATGAGGCAACGTGTTATGATGATGGGAAGACATACCACGTAGGAGAACAGTGGCAGAAGGAATATCTCGGTGCCATTTGCTCCTGCACATGCTTTGGAGGCCAGCGGGGCTGGCGCTGTGACAACTGCCGCAGACCTGGGGGTGAACCCAGTCCCGAAGGCACTACTGGCCAGTCCTACAACCAGTATTCTCAGAGATACCATCAGAGAACAAACACTAATGTTAATTGCCCAATTGAGTGCTTCATGCCTTTAGATGTACAGGCTGACAGAGAAGATTCCCGAGAGTAA，

为野生型FN1基因编码DNA参考序列的第2372位突变前碱基。

c.2372A>G表示：与野生型FN1基因编码DNA的参考序列相比，突变基因FN1的第2372位点的碱基A突变为碱基G，具体编码DNA的核苷酸序列为SEQ ID NO:3。

(4)转录本号为NM_001306129时，野生型FN1基因编码蛋白为：

MLRGPGPGLLLLAVQCLGTAVPSTGASKSKRQAQQMVQPQSPVAVSQSKPGCYDNGKHYQINQQWERTYLGNALVCTCYGGSRGFNCESKPEAEETCFDKYTGNTYRVGDTYERPKDSMIWDCTCIGAGRGRISCTIANRCHEGGQSYKIGDTWRRPHETGGYMLECVCLGNGKGEWTCKPIAEKCFDHAAGTSYVVGETWEKPYQGWMMVDCTCLGEGSGRITCTSRNRCNDQDTRTSYRIGDTWSKKDNRGNLLQCICTGNGRGEWKCERHTSVQTTSSGSGPFTDVRAAVYQPQPHPQPPPYGHCVTDSGVVYSVGMQWLKTQGNKQMLCTCLGNGVSCQETAVTQTYGGNSNGEPCVLPFTYNGRTFYSCTTEGRQDGHLWCSTTSNYEQDQKYSFCTDHTVLVQTRGGNSNGALCHFPFLYNNHNYTDCTSEGRRDNMKWCGTTQNYDADQKFGFCPMAAHEEICTTNEGVMYRIGDQWDKQHDMGHMMRCTCVGNGRGEWTCIAYSQLRDQCIVDDITYNVNDTFHKRHEEGHMLNCTCFGQGRGRWKCDPVDQCQDSETGTFYQIGDSWEKYVHGVRYQCYCYGRGIGEWHCQPLQTYPSSSGPVEVFITETPSQPNSHPIQWNAPQPSHISKYILRWRPKNSVGRWKEATIPGHLNSYTIKGLKPGVVYEGQLISIQQYGHQEVTRFDFTTTSTSTPVTSNTVTGETTPFSPLVATSESVTEITASSFVVSWVSASDTVSGFRVEYELSEEGDEPQYLDLPSTATSVNIPDLLPGRKYIVNV

QISEDGEQSLILSTSQTTAPDAPPDTTVDQVDDTSIVVRWSRPQAPITGYRIVYSPSVEGSSTELNLPETANSVTLSDLQPGVQYNITIYAVEENQESTPVVIQQETTGTPRSDTVPSPRDLQFVEVTDVKVTIMWTPPESAVTGYRVDVIPVNLPGEHGQRLPISRNTFAEVTGLSPGVTYYFKVFAVSHGRESKPLTAQQTTKLDAPTNLQFVNETDSTVLVRWTPPRAQITGYRLTVGLTRRGQPRQYNVGPSVSKYPLRNLQPASEYTVSLVAIKGNQESPKATGVFTTLQPGSSIPPYNTEVTETTIVITWTPAPRIGFKLGVRPSQGGEAPREVTSDSGSIVVSGLTPGVEYVYTIQVLRDGQERDAPIVNKVVTPLSPPTNLHLEANPDTGVLTVSWERSTTPDITGYRITTTPTNGQQGNSLEEVVHADQSSCTFDNLSPGLEYNVSVYTVKDDKESVPISDTIIPEVPQLTDLSFVDITDSSIGLRWTPLNSSTIIGYRITVVAAGEGIPIFEDFVDSSVGYYTVTGLEPGIDYDISVITLINGGESAPTTLTQQTAVPPPTDLRFTNIGPDTMRVTWAPPPSIDLTNFLVRYSPVKNEEDVAELSISPSDNAVVLTNLLPGTEYVVSVSSVYEQHESTPLRGRQKTGLDSPTGIDFSDITANSFTVHWIAPRATITGYRIRHHPEHFSGRPREDRVPHSRNSITLTNLTPGTEYVVSIVALNGREESPLLIGQQSTVSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKSTATISGLKPGVDYTITVYAVTGRGDSPASSKPISINYRTEIDKPSQMQVTDVQDNSISVKWLPSSSPVTGYRVTTTPKNGPGPTKTKTAGPDQTEMTIEGLQPTVEYVVSVYAQNPSGESQPLVQTAVTNIDRPKGLAFTDVDVDSIKIAWESPQGQVSRYRVTYSSPEDGIHELFPAPDGEEDTAELQGLRPGSEYTVSVVALHDDMESQPLIGTQSTAIPAPTDLKFTQVTPTSLSAQWTPPNVQLTGYRVRVTPKEKTGPMKEINLAPDSSSVVVSGLMVATKYEVSVYALKDTLTSRPAQGVVTTLENVSPPRRARVTDATETTITISWRTKTETITGFQVDAVPANGQTPIQRTIKPDVRSYTITGLQPGTDYKIYLYTLNDNARSSPVVIDASTAIDAPSNLRFLATTPNSLLVSWQPPRARITGYIIKYEKPGSPPREVVPRPRPGVTEATITGLEPGTEYTIYVIALKNNQKSEPLIGRKKTDELPQLVTLPHPNLHGPEILDVPSTVQKTPFVTHPGYDTGNGIQLPGTSGQQPSVGQQMIFEEHGFRRTTPPTTATPIRHRPRPYPPNVGQEALSQTTISWAPFQDTSEYIISCHPVGTDEEPLQFRVPGTSTSATLTGLTRGATYNVIVEALKDQQRHKVREEVVTVGNSVNEGLNQPTDDSCFDPYTVSHYAVGDEWERMSESGFKLLCQCLGFGSGHFRCDSSRWCHDNGVNYKIGEKWDRQGENGQMMSCTCLGNGKGEFKCDPHEATCYDDGKTYHVGEQWQKEYLGAICSCTCFGGQRGWRCDNCRRPGGEPSPEGTTGQSYNQYSQRYHQRTNTNVNCPIECFMPLDVQADREDSRE。

为野生型FN1基因编码蛋白的第791位酪氨酸(Tyr,Y)。

p.Tyr791Cys表示：与野生型FN1基因编码蛋白的氨基酸序列相比，突变基因FN1编码蛋白的氨基酸第791位的酪氨酸(Tyr,Y)突变为半胱氨酸(Cys,C)，具体氨基酸序列为SEQID NO:4。

(5)查询人群频率数据库发现FN1基因c.2372A>G杂合错义变异(FN1:p.Tyr791Cyshet)为罕见变异(千人基因组：无，ESP6500：无，ExAC：无)。

采用多个生物信息预测软件(包括SIFT和Polyphen-2等)交叉预测，结果多为有害(SIFT为“D”，Polyphen-2为“D”，MutationTaster_pred为“D”，VEST3评分为“0.882”，其他为“3个T/2个D/1个M”)，氨基酸由极性不带电荷的酪氨酸变为极性不带电荷的半胱氨酸，提示该变异所致的氨基酸改变可能会对蛋白功能产生影响。查询数据库发现该位置的氨基酸在脊椎动物中保守性好。查询ClinVar、HGMD数据库未发现该变异，文献检索未发现该变异与疾病相关的报导。根据现有证据：该变异为罕见变异、软件预测该变异可能会对蛋白功能产生影响、该位置氨基酸在脊椎动物中保守性好，但缺乏家系连锁及功能学证据支持，所以该变异为可疑致病变异(C1级)。

实施例2-与遗传性肾小球疾病相关的突变基因的检测试剂盒

与遗传性肾小球疾病相关的突变基因的检测试剂盒，包括Taq DNA聚合酶、PCR缓冲液和引物等。具体引物如下：

上游引物(FN1-E16F，SEQ ID NO:1)：5'AGCACATTACCTTCTAGTCGCTT 3”；

下游引物(FN1-E16R，SEQ ID NO:2)：5'AACTTGGTCCACAGTCGTGTC 3'；

长度：394bp。

利用本试剂盒筛选突变的致病基因FN1的具体步骤为：提取待测者DNA，然后使用经设计的引物组合(SEQ ID NO:1和SEQ ID NO:2)对FN1基因进行扩增，得到PCR产物，使用1.5％的琼脂糖凝胶电泳检测PCR产物，选用1000bp Marker作为参考，检测验证扩增产物为预期的大小，最后对PCR产物进行测序。从NCBI(https://www.ncbi.nlm.nih.gov/)数据库获得参考序列和测序结果进行比对，判断待测者FN1基因是否携带c.2372A>G杂合错义变异，协助临床确诊纤维连接蛋白肾小球病。

实施例3-家系验证实验

本实施例采用家系连锁分析方法验证与遗传性肾小球疾病相关的突变基因的致病性。

具体地，选取一个家族性纤维连接蛋白肾小球病家系中的三代成员，该家系中的先证者(女，15岁)临床被诊断为纤维连接蛋白肾小球病。

在先证者及其家属自愿签署知情同意书的前提下，寄送5-10mL全血样本，建立病历资料库，详细记录先证者病情、家系情况等资料。本研究已得到本单位伦理委员会批准。

先证者临床概况描述：

表3先证者临床概况

采用实施例2提供的检测试剂盒对先证者及其家属的FN1基因进行基因检测，结果如图1-图3所示，图1为实施例3的家系图；图2为实施例3中先证者、先证者母亲等人的Sanger测序图；图3为实施例3家系中先证者父亲、先证者妹妹等人的Sanger测序图。

如图1-图3所示，家系中临床诊断出纤维连接蛋白肾小球病的先证者、先证者母亲等人均携带了突变的FN1基因，而未患有纤维连接蛋白肾小球病的家系成员均未携带突变的FN1基因，由此验证可以说明突变的FN1基因对纤维连接蛋白肾小球病的致病性。

实施例4-针对家系外正常人的验证实验

采用实施例2的检测试剂盒，对700例家系外正常人进行FN1基因检测，结果均未能检测到该突变。

实施例5-针对家系外家族遗传性纤维连接蛋白肾小球病患者的验证实验

在中国范围内，对患有纤维连接蛋白肾小球病、肥厚型心肌病、扩张型心肌病、长QT等单基因遗传病的患者中检测FN1基因，患者总共1900例，每种疾病的患病人数不等，结果显示，除实施例3提供的家系外，仅在临床诊断患有纤维连接蛋白肾小球病的2例患者中检测到具有c.2372A>G杂合错义变异的突变基因FN1。

本实验再次验证说明突变的FN1致病基因的会导致纤维连接蛋白肾小球病，支持临床诊断。

以上详细描述了本发明的较佳具体实施例。应当理解，本领域的普通技术人员无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此，凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案，皆应在由权利要求书所确定的保护范围内。

序列表

<110> 百世诺（北京）医学检验实验室有限公司

<120> 与遗传性肾小球疾病相关的突变基因及其应用

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atgcttaggg gtccggggcc cgggctgctg ctgctggccg tccagtgcct ggggacagcg 60

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ataaatcaac agtgggagcg gacctaccta ggcaatgcgt tggtttgtac ttgttatgga 240

ggaagccgag gttttaactg cgagagtaaa cctgaagctg aagagacttg ctttgacaag 300

tacactggga acacttaccg agtgggtgac acttatgagc gtcctaaaga ctccatgatc 360

tgggactgta cctgcatcgg ggctgggcga gggagaataa gctgtaccat cgcaaaccgc 420

tgccatgaag ggggtcagtc ctacaagatt ggtgacacct ggaggagacc acatgagact 480

ggtggttaca tgttagagtg tgtgtgtctt ggtaatggaa aaggagaatg gacctgcaag 540

cccatagctg agaagtgttt tgatcatgct gctgggactt cctatgtggt cggagaaacg 600

tgggagaagc cctaccaagg ctggatgatg gtagattgta cttgcctggg agaaggcagc 660

ggacgcatca cttgcacttc tagaaataga tgcaacgatc aggacacaag gacatcctat 720

agaattggag acacctggag caagaaggat aatcgaggaa acctgctcca gtgcatctgc 780

acaggcaacg gccgaggaga gtggaagtgt gagaggcaca cctctgtgca gaccacatcg 840

agcggatctg gccccttcac cgatgttcgt gcagctgttt accaaccgca gcctcacccc 900

cagcctcctc cctatggcca ctgtgtcaca gacagtggtg tggtctactc tgtggggatg 960

cagtggctga agacacaagg aaataagcaa atgctttgca cgtgcctggg caacggagtc 1020

agctgccaag agacagctgt aacccagact tacggtggca actcaaatgg agagccatgt 1080

gtcttaccat tcacctacaa tggcaggacg ttctactcct gcaccacaga agggcgacag 1140

gacggacatc tttggtgcag cacaacttcg aattatgagc aggaccagaa atactctttc 1200

tgcacagacc acactgtttt ggttcagact cgaggaggaa attccaatgg tgccttgtgc 1260

cacttcccct tcctatacaa caaccacaat tacactgatt gcacttctga gggcagaaga 1320

gacaacatga agtggtgtgg gaccacacag aactatgatg ccgaccagaa gtttgggttc 1380

tgccccatgg ctgcccacga ggaaatctgc acaaccaatg aaggggtcat gtaccgcatt 1440

ggagatcagt gggataagca gcatgacatg ggtcacatga tgaggtgcac gtgtgttggg 1500

aatggtcgtg gggaatggac atgcattgcc tactcgcagc ttcgagatca gtgcattgtt 1560

gatgacatca cttacaatgt gaacgacaca ttccacaagc gtcatgaaga ggggcacatg 1620

ctgaactgta catgcttcgg tcagggtcgg ggcaggtgga agtgtgatcc cgtcgaccaa 1680

tgccaggatt cagagactgg gacgttttat caaattggag attcatggga gaagtatgtg 1740

catggtgtca gataccagtg ctactgctat ggccgtggca ttggggagtg gcattgccaa 1800

cctttacaga cctatccaag ctcaagtggt cctgtcgaag tatttatcac tgagactccg 1860

agtcagccca actcccaccc catccagtgg aatgcaccac agccatctca catttccaag 1920

tacattctca ggtggagacc taaaaattct gtaggccgtt ggaaggaagc taccatacca 1980

ggccacttaa actcctacac catcaaaggc ctgaagcctg gtgtggtata cgagggccag 2040

ctcatcagca tccagcagta cggccaccaa gaagtgactc gctttgactt caccaccacc 2100

agcaccagca cacctgtgac cagcaacacc gtgacaggag agacgactcc cttttctcct 2160

cttgtggcca cttctgaatc tgtgaccgaa atcacagcca gtagctttgt ggtctcctgg 2220

gtctcagctt ccgacaccgt gtcgggattc cgggtggaat atgagctgag tgaggaggga 2280

gatgagccac agtacctgga tcttccaagc acagccactt ctgtgaacat ccctgacctg 2340

cttcctggcc gaaaatacat tgtaaatgtc tgtcagatat ctgaggatgg ggagcagagt 2400

ttgatcctgt ctacttcaca aacaacagcg cctgatgccc ctcctgacac gactgtggac 2460

caagttgatg acacctcaat tgttgttcgc tggagcagac cccaggctcc catcacaggg 2520

tacagaatag tctattcgcc atcagtagaa ggtagcagca cagaactcaa ccttcctgaa 2580

actgcaaact ccgtcaccct cagtgacttg caacctggtg ttcagtataa catcactatc 2640

tatgctgtgg aagaaaatca agaaagtaca cctgttgtca ttcaacaaga aaccactggc 2700

accccacgct cagatacagt gccctctccc agggacctgc agtttgtgga agtgacagac 2760

gtgaaggtca ccatcatgtg gacaccgcct gagagtgcag tgaccggcta ccgtgtggat 2820

gtgatccccg tcaacctgcc tggcgagcac gggcagaggc tgcccatcag caggaacacc 2880

tttgcagaag tcaccgggct gtcccctggg gtcacctatt acttcaaagt ctttgcagtg 2940

agccatggga gggagagcaa gcctctgact gctcaacaga caaccaaact ggatgctccc 3000

actaacctcc agtttgtcaa tgaaactgat tctactgtcc tggtgagatg gactccacct 3060

cgggcccaga taacaggata ccgactgacc gtgggcctta cccgaagagg acagcccagg 3120

cagtacaatg tgggtccctc tgtctccaag tacccactga ggaatctgca gcctgcatct 3180

gagtacaccg tatccctcgt ggccataaag ggcaaccaag agagccccaa agccactgga 3240

gtctttacca cactgcagcc tgggagctct attccacctt acaacaccga ggtgactgag 3300

accaccattg tgatcacatg gacgcctgct ccaagaattg gttttaagct gggtgtacga 3360

ccaagccagg gaggagaggc accacgagaa gtgacttcag actcaggaag catcgttgtg 3420

tccggcttga ctccaggagt agaatacgtc tacaccatcc aagtcctgag agatggacag 3480

gaaagagatg cgccaattgt aaacaaagtg gtgacaccat tgtctccacc aacaaacttg 3540

catctggagg caaaccctga cactggagtg ctcacagtct cctgggagag gagcaccacc 3600

ccagacatta ctggttatag aattaccaca acccctacaa acggccagca gggaaattct 3660

ttggaagaag tggtccatgc tgatcagagc tcctgcactt ttgataacct gagtcccggc 3720

ctggagtaca atgtcagtgt ttacactgtc aaggatgaca aggaaagtgt ccctatctct 3780

gataccatca tcccagaggt gccccaactc actgacctaa gctttgttga tataaccgat 3840

tcaagcatcg gcctgaggtg gaccccgcta aactcttcca ccattattgg gtaccgcatc 3900

acagtagttg cggcaggaga aggtatccct atttttgaag attttgtgga ctcctcagta 3960

ggatactaca cagtcacagg gctggagccg ggcattgact atgatatcag cgttatcact 4020

ctcattaatg gcggcgagag tgcccctact acactgacac aacaaacggc tgttcctcct 4080

cccactgacc tgcgattcac caacattggt ccagacacca tgcgtgtcac ctgggctcca 4140

cccccatcca ttgatttaac caacttcctg gtgcgttact cacctgtgaa aaatgaggaa 4200

gatgttgcag agttgtcaat ttctccttca gacaatgcag tggtcttaac aaatctcctg 4260

cctggtacag aatatgtagt gagtgtctcc agtgtctacg aacaacatga gagcacacct 4320

cttagaggaa gacagaaaac aggtcttgat tccccaactg gcattgactt ttctgatatt 4380

actgccaact cttttactgt gcactggatt gctcctcgag ccaccatcac tggctacagg 4440

atccgccatc atcccgagca cttcagtggg agacctcgag aagatcgggt gccccactct 4500

cggaattcca tcaccctcac caacctcact ccaggcacag agtatgtggt cagcatcgtt 4560

gctcttaatg gcagagagga aagtccctta ttgattggcc aacaatcaac agtttctgat 4620

gttccgaggg acctggaagt tgttgctgcg acccccacca gcctactgat cagctgggat 4680

gctcctgctg tcacagtgag atattacagg atcacttacg gagagacagg aggaaatagc 4740

cctgtccagg agttcactgt gcctgggagc aagtctacag ctaccatcag cggccttaaa 4800

cctggagttg attataccat cactgtgtat gctgtcactg gccgtggaga cagccccgca 4860

agcagcaagc caatttccat taattaccga acagaaattg acaaaccatc ccagatgcaa 4920

gtgaccgatg ttcaggacaa cagcattagt gtcaagtggc tgccttcaag ttcccctgtt 4980

actggttaca gagtaaccac cactcccaaa aatggaccag gaccaacaaa aactaaaact 5040

gcaggtccag atcaaacaga aatgactatt gaaggcttgc agcccacagt ggagtatgtg 5100

gttagtgtct atgctcagaa tccaagcgga gagagtcagc ctctggttca gactgcagta 5160

accaacattg atcgccctaa aggactggca ttcactgatg tggatgtcga ttccatcaaa 5220

attgcttggg aaagcccaca ggggcaagtt tccaggtaca gggtgaccta ctcgagccct 5280

gaggatggaa tccatgagct attccctgca cctgatggtg aagaagacac tgcagagctg 5340

caaggcctca gaccgggttc tgagtacaca gtcagtgtgg ttgccttgca cgatgatatg 5400

gagagccagc ccctgattgg aacccagtcc acagctattc ctgcaccaac tgacctgaag 5460

ttcactcagg tcacacccac aagcctgagc gcccagtgga caccacccaa tgttcagctc 5520

actggatatc gagtgcgggt gacccccaag gagaagaccg gaccaatgaa agaaatcaac 5580

cttgctcctg acagctcatc cgtggttgta tcaggactta tggtggccac caaatatgaa 5640

gtgagtgtct atgctcttaa ggacactttg acaagcagac cagctcaggg agttgtcacc 5700

actctggaga atgtcagccc accaagaagg gctcgtgtga cagatgctac tgagaccacc 5760

atcaccatta gctggagaac caagactgag acgatcactg gcttccaagt tgatgccgtt 5820

ccagccaatg gccagactcc aatccagaga accatcaagc cagatgtcag aagctacacc 5880

atcacaggtt tacaaccagg cactgactac aagatctacc tgtacacctt gaatgacaat 5940

gctcggagct cccctgtggt catcgacgcc tccactgcca ttgatgcacc atccaacctg 6000

cgtttcctgg ccaccacacc caattccttg ctggtatcat ggcagccgcc acgtgccagg 6060

attaccggct acatcatcaa gtatgagaag cctgggtctc ctcccagaga agtggtccct 6120

cggccccgcc ctggtgtcac agaggctact attactggcc tggaaccggg aaccgaatat 6180

acaatttatg tcattgccct gaagaataat cagaagagcg agcccctgat tggaaggaaa 6240

aagacagacg agcttcccca actggtaacc cttccacacc ccaatcttca tggaccagag 6300

atcttggatg ttccttccac agttcaaaag acccctttcg tcacccaccc tgggtatgac 6360

actggaaatg gtattcagct tcctggcact tctggtcagc aacccagtgt tgggcaacaa 6420

atgatctttg aggaacatgg ttttaggcgg accacaccgc ccacaacggc cacccccata 6480

aggcataggc caagaccata cccgccgaat gtaggacaag aagctctctc tcagacaacc 6540

atctcatggg ccccattcca ggacacttct gagtacatca tttcatgtca tcctgttggc 6600

actgatgaag aacccttaca gttcagggtt cctggaactt ctaccagtgc cactctgaca 6660

ggcctcacca gaggtgccac ctacaacgtc atagtggagg cactgaaaga ccagcagagg 6720

cataaggttc gggaagaggt tgttaccgtg ggcaactctg tcaacgaagg cttgaaccaa 6780

cctacggatg actcgtgctt tgacccctac acagtttccc attatgccgt tggagatgag 6840

tgggaacgaa tgtctgaatc aggctttaaa ctgttgtgcc agtgcttagg ctttggaagt 6900

ggtcatttca gatgtgattc atctagatgg tgccatgaca atggtgtgaa ctacaagatt 6960

ggagagaagt gggaccgtca gggagaaaat ggccagatga tgagctgcac atgtcttggg 7020

aacggaaaag gagaattcaa gtgtgaccct catgaggcaa cgtgttatga tgatgggaag 7080

acataccacg taggagaaca gtggcagaag gaatatctcg gtgccatttg ctcctgcaca 7140

tgctttggag gccagcgggg ctggcgctgt gacaactgcc gcagacctgg gggtgaaccc 7200

agtcccgaag gcactactgg ccagtcctac aaccagtatt ctcagagata ccatcagaga 7260

acaaacacta atgttaattg cccaattgag tgcttcatgc ctttagatgt acaggctgac 7320

agagaagatt cccgagagta a 7341

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

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Ala Gln Gln Met Val Gln Pro Gln Ser Pro Val Ala Val Ser Gln Ser

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Trp Glu Arg Thr Tyr Leu Gly Asn Ala Leu Val Cys Thr Cys Tyr Gly

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Gly Ser Arg Gly Phe Asn Cys Glu Ser Lys Pro Glu Ala Glu Glu Thr

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Cys Phe Asp Lys Tyr Thr Gly Asn Thr Tyr Arg Val Gly Asp Thr Tyr

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Glu Arg Pro Lys Asp Ser Met Ile Trp Asp Cys Thr Cys Ile Gly Ala

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Gly Arg Gly Arg Ile Ser Cys Thr Ile Ala Asn Arg Cys His Glu Gly

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Gly Gln Ser Tyr Lys Ile Gly Asp Thr Trp Arg Arg Pro His Glu Thr

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Gly Gly Tyr Met Leu Glu Cys Val Cys Leu Gly Asn Gly Lys Gly Glu

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Trp Thr Cys Lys Pro Ile Ala Glu Lys Cys Phe Asp His Ala Ala Gly

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Thr Ser Tyr Val Val Gly Glu Thr Trp Glu Lys Pro Tyr Gln Gly Trp

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Met Met Val Asp Cys Thr Cys Leu Gly Glu Gly Ser Gly Arg Ile Thr

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Cys Thr Ser Arg Asn Arg Cys Asn Asp Gln Asp Thr Arg Thr Ser Tyr

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Arg Ile Gly Asp Thr Trp Ser Lys Lys Asp Asn Arg Gly Asn Leu Leu

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Gln Cys Ile Cys Thr Gly Asn Gly Arg Gly Glu Trp Lys Cys Glu Arg

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His Thr Ser Val Gln Thr Thr Ser Ser Gly Ser Gly Pro Phe Thr Asp

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Val Arg Ala Ala Val Tyr Gln Pro Gln Pro His Pro Gln Pro Pro Pro

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Tyr Gly His Cys Val Thr Asp Ser Gly Val Val Tyr Ser Val Gly Met

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Gln Trp Leu Lys Thr Gln Gly Asn Lys Gln Met Leu Cys Thr Cys Leu

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Gly Asn Gly Val Ser Cys Gln Glu Thr Ala Val Thr Gln Thr Tyr Gly

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Gly Asn Ser Asn Gly Glu Pro Cys Val Leu Pro Phe Thr Tyr Asn Gly

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Arg Thr Phe Tyr Ser Cys Thr Thr Glu Gly Arg Gln Asp Gly His Leu

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Trp Cys Ser Thr Thr Ser Asn Tyr Glu Gln Asp Gln Lys Tyr Ser Phe

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Cys Thr Asp His Thr Val Leu Val Gln Thr Arg Gly Gly Asn Ser Asn

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Gly Ala Leu Cys His Phe Pro Phe Leu Tyr Asn Asn His Asn Tyr Thr

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Asp Cys Thr Ser Glu Gly Arg Arg Asp Asn Met Lys Trp Cys Gly Thr

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Thr Gln Asn Tyr Asp Ala Asp Gln Lys Phe Gly Phe Cys Pro Met Ala

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Ala His Glu Glu Ile Cys Thr Thr Asn Glu Gly Val Met Tyr Arg Ile

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Gly Asp Gln Trp Asp Lys Gln His Asp Met Gly His Met Met Arg Cys

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Thr Cys Val Gly Asn Gly Arg Gly Glu Trp Thr Cys Ile Ala Tyr Ser

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Gln Leu Arg Asp Gln Cys Ile Val Asp Asp Ile Thr Tyr Asn Val Asn

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Asp Thr Phe His Lys Arg His Glu Glu Gly His Met Leu Asn Cys Thr

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Cys Phe Gly Gln Gly Arg Gly Arg Trp Lys Cys Asp Pro Val Asp Gln

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Cys Gln Asp Ser Glu Thr Gly Thr Phe Tyr Gln Ile Gly Asp Ser Trp

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Glu Lys Tyr Val His Gly Val Arg Tyr Gln Cys Tyr Cys Tyr Gly Arg

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

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

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Ser His Pro Ile Gln Trp Asn Ala Pro Gln Pro Ser His Ile Ser Lys

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Tyr Ile Leu Arg Trp Arg Pro Lys Asn Ser Val Gly Arg Trp Lys Glu

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Ala Thr Ile Pro Gly His Leu Asn Ser Tyr Thr Ile Lys Gly Leu Lys

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

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His Gln Glu Val Thr Arg Phe Asp Phe Thr Thr Thr Ser Thr Ser Thr

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Pro Val Thr Ser Asn Thr Val Thr Gly Glu Thr Thr Pro Phe Ser Pro

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Leu Val Ala Thr Ser Glu Ser Val Thr Glu Ile Thr Ala Ser Ser Phe

725 730 735

Val Val Ser Trp Val Ser Ala Ser Asp Thr Val Ser Gly Phe Arg Val

740 745 750

Glu Tyr Glu Leu Ser Glu Glu Gly Asp Glu Pro Gln Tyr Leu Asp Leu

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Pro Ser Thr Ala Thr Ser Val Asn Ile Pro Asp Leu Leu Pro Gly Arg

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Lys Tyr Ile Val Asn Val Cys Gln Ile Ser Glu Asp Gly Glu Gln Ser

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Leu Ile Leu Ser Thr Ser Gln Thr Thr Ala Pro Asp Ala Pro Pro Asp

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Thr Thr Val Asp Gln Val Asp Asp Thr Ser Ile Val Val Arg Trp Ser

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

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Val Glu Gly Ser Ser Thr Glu Leu Asn Leu Pro Glu Thr Ala Asn Ser

850 855 860

Val Thr Leu Ser Asp Leu Gln Pro Gly Val Gln Tyr Asn Ile Thr Ile

865 870 875 880

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

885 890 895

Glu Thr Thr Gly Thr Pro Arg Ser Asp Thr Val Pro Ser Pro Arg Asp

900 905 910

Leu Gln Phe Val Glu Val Thr Asp Val Lys Val Thr Ile Met Trp Thr

915 920 925

Pro Pro Glu Ser Ala Val Thr Gly Tyr Arg Val Asp Val Ile Pro Val

930 935 940

Asn Leu Pro Gly Glu His Gly Gln Arg Leu Pro Ile Ser Arg Asn Thr

945 950 955 960

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

965 970 975

Val Phe Ala Val Ser His Gly Arg Glu Ser Lys Pro Leu Thr Ala Gln

980 985 990

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

995 1000 1005

Thr Asp Ser Thr Val Leu Val Arg Trp Thr Pro Pro Arg Ala Gln Ile

1010 1015 1020

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

1025 1030 1035 1040

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

1045 1050 1055

Gln Pro Ala Ser Glu Tyr Thr Val Ser Leu Val Ala Ile Lys Gly Asn

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

1075 1080 1085

Ser Ser Ile Pro Pro Tyr Asn Thr Glu Val Thr Glu Thr Thr Ile Val

1090 1095 1100

Ile Thr Trp Thr Pro Ala Pro Arg Ile Gly Phe Lys Leu Gly Val Arg

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Pro Ser Gln Gly Gly Glu Ala Pro Arg Glu Val Thr Ser Asp Ser Gly

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Ser Ile Val Val Ser Gly Leu Thr Pro Gly Val Glu Tyr Val Tyr Thr

1140 1145 1150

Ile Gln Val Leu Arg Asp Gly Gln Glu Arg Asp Ala Pro Ile Val Asn

1155 1160 1165

Lys Val Val Thr Pro Leu Ser Pro Pro Thr Asn Leu His Leu Glu Ala

1170 1175 1180

Asn Pro Asp Thr Gly Val Leu Thr Val Ser Trp Glu Arg Ser Thr Thr

1185 1190 1195 1200

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

1205 1210 1215

Gln Gly Asn Ser Leu Glu Glu Val Val His Ala Asp Gln Ser Ser Cys

1220 1225 1230

Thr Phe Asp Asn Leu Ser Pro Gly Leu Glu Tyr Asn Val Ser Val Tyr

1235 1240 1245

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

1250 1255 1260

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

1265 1270 1275 1280

Ser Ser Ile Gly Leu Arg Trp Thr Pro Leu Asn Ser Ser Thr Ile Ile

1285 1290 1295

Gly Tyr Arg Ile Thr Val Val Ala Ala Gly Glu Gly Ile Pro Ile Phe

1300 1305 1310

Glu Asp Phe Val Asp Ser Ser Val Gly Tyr Tyr Thr Val Thr Gly Leu

1315 1320 1325

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

1330 1335 1340

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

1345 1350 1355 1360

Pro Thr Asp Leu Arg Phe Thr Asn Ile Gly Pro Asp Thr Met Arg Val

1365 1370 1375

Thr Trp Ala Pro Pro Pro Ser Ile Asp Leu Thr Asn Phe Leu Val Arg

1380 1385 1390

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

1395 1400 1405

Pro Ser Asp Asn Ala Val Val Leu Thr Asn Leu Leu Pro Gly Thr Glu

1410 1415 1420

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

1425 1430 1435 1440

Leu Arg Gly Arg Gln Lys Thr Gly Leu Asp Ser Pro Thr Gly Ile Asp

1445 1450 1455

Phe Ser Asp Ile Thr Ala Asn Ser Phe Thr Val His Trp Ile Ala Pro

1460 1465 1470

Arg Ala Thr Ile Thr Gly Tyr Arg Ile Arg His His Pro Glu His Phe

1475 1480 1485

Ser Gly Arg Pro Arg Glu Asp Arg Val Pro His Ser Arg Asn Ser Ile

1490 1495 1500

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

1505 1510 1515 1520

Ala Leu Asn Gly Arg Glu Glu Ser Pro Leu Leu Ile Gly Gln Gln Ser

1525 1530 1535

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

1540 1545 1550

Thr Ser Leu Leu Ile Ser Trp Asp Ala Pro Ala Val Thr Val Arg Tyr

1555 1560 1565

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

1570 1575 1580

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

1585 1590 1595 1600

Pro Gly Val Asp Tyr Thr Ile Thr Val Tyr Ala Val Thr Gly Arg Gly

1605 1610 1615

Asp Ser Pro Ala Ser Ser Lys Pro Ile Ser Ile Asn Tyr Arg Thr Glu

1620 1625 1630

Ile Asp Lys Pro Ser Gln Met Gln Val Thr Asp Val Gln Asp Asn Ser

1635 1640 1645

Ile Ser Val Lys Trp Leu Pro Ser Ser Ser Pro Val Thr Gly Tyr Arg

1650 1655 1660

Val Thr Thr Thr Pro Lys Asn Gly Pro Gly Pro Thr Lys Thr Lys Thr

1665 1670 1675 1680

Ala Gly Pro Asp Gln Thr Glu Met Thr Ile Glu Gly Leu Gln Pro Thr

1685 1690 1695

Val Glu Tyr Val Val Ser Val Tyr Ala Gln Asn Pro Ser Gly Glu Ser

1700 1705 1710

Gln Pro Leu Val Gln Thr Ala Val Thr Asn Ile Asp Arg Pro Lys Gly

1715 1720 1725

Leu Ala Phe Thr Asp Val Asp Val Asp Ser Ile Lys Ile Ala Trp Glu

1730 1735 1740

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

1745 1750 1755 1760

Glu Asp Gly Ile His Glu Leu Phe Pro Ala Pro Asp Gly Glu Glu Asp

1765 1770 1775

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

1780 1785 1790

Val Val Ala Leu His Asp Asp Met Glu Ser Gln Pro Leu Ile Gly Thr

1795 1800 1805

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

1810 1815 1820

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

1825 1830 1835 1840

Thr Gly Tyr Arg Val Arg Val Thr Pro Lys Glu Lys Thr Gly Pro Met

1845 1850 1855

Lys Glu Ile Asn Leu Ala Pro Asp Ser Ser Ser Val Val Val Ser Gly

1860 1865 1870

Leu Met Val Ala Thr Lys Tyr Glu Val Ser Val Tyr Ala Leu Lys Asp

1875 1880 1885

Thr Leu Thr Ser Arg Pro Ala Gln Gly Val Val Thr Thr Leu Glu Asn

1890 1895 1900

Val Ser Pro Pro Arg Arg Ala Arg Val Thr Asp Ala Thr Glu Thr Thr

1905 1910 1915 1920

Ile Thr Ile Ser Trp Arg Thr Lys Thr Glu Thr Ile Thr Gly Phe Gln

1925 1930 1935

Val Asp Ala Val Pro Ala Asn Gly Gln Thr Pro Ile Gln Arg Thr Ile

1940 1945 1950

Lys Pro Asp Val Arg Ser Tyr Thr Ile Thr Gly Leu Gln Pro Gly Thr

1955 1960 1965

Asp Tyr Lys Ile Tyr Leu Tyr Thr Leu Asn Asp Asn Ala Arg Ser Ser

1970 1975 1980

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

1985 1990 1995 2000

Arg Phe Leu Ala Thr Thr Pro Asn Ser Leu Leu Val Ser Trp Gln Pro

2005 2010 2015

Pro Arg Ala Arg Ile Thr Gly Tyr Ile Ile Lys Tyr Glu Lys Pro Gly

2020 2025 2030

Ser Pro Pro Arg Glu Val Val Pro Arg Pro Arg Pro Gly Val Thr Glu

2035 2040 2045

Ala Thr Ile Thr Gly Leu Glu Pro Gly Thr Glu Tyr Thr Ile Tyr Val

2050 2055 2060

Ile Ala Leu Lys Asn Asn Gln Lys Ser Glu Pro Leu Ile Gly Arg Lys

2065 2070 2075 2080

Lys Thr Asp Glu Leu Pro Gln Leu Val Thr Leu Pro His Pro Asn Leu

2085 2090 2095

His Gly Pro Glu Ile Leu Asp Val Pro Ser Thr Val Gln Lys Thr Pro

2100 2105 2110

Phe Val Thr His Pro Gly Tyr Asp Thr Gly Asn Gly Ile Gln Leu Pro

2115 2120 2125

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

2130 2135 2140

Glu His Gly Phe Arg Arg Thr Thr Pro Pro Thr Thr Ala Thr Pro Ile

2145 2150 2155 2160

Arg His Arg Pro Arg Pro Tyr Pro Pro Asn Val Gly Gln Glu Ala Leu

2165 2170 2175

Ser Gln Thr Thr Ile Ser Trp Ala Pro Phe Gln Asp Thr Ser Glu Tyr

2180 2185 2190

Ile Ile Ser Cys His Pro Val Gly Thr Asp Glu Glu Pro Leu Gln Phe

2195 2200 2205

Arg Val Pro Gly Thr Ser Thr Ser Ala Thr Leu Thr Gly Leu Thr Arg

2210 2215 2220

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

2225 2230 2235 2240

His Lys Val Arg Glu Glu Val Val Thr Val Gly Asn Ser Val Asn Glu

2245 2250 2255

Gly Leu Asn Gln Pro Thr Asp Asp Ser Cys Phe Asp Pro Tyr Thr Val

2260 2265 2270

Ser His Tyr Ala Val Gly Asp Glu Trp Glu Arg Met Ser Glu Ser Gly

2275 2280 2285

Phe Lys Leu Leu Cys Gln Cys Leu Gly Phe Gly Ser Gly His Phe Arg

2290 2295 2300

Cys Asp Ser Ser Arg Trp Cys His Asp Asn Gly Val Asn Tyr Lys Ile

2305 2310 2315 2320

Gly Glu Lys Trp Asp Arg Gln Gly Glu Asn Gly Gln Met Met Ser Cys

2325 2330 2335

Thr Cys Leu Gly Asn Gly Lys Gly Glu Phe Lys Cys Asp Pro His Glu

2340 2345 2350

Ala Thr Cys Tyr Asp Asp Gly Lys Thr Tyr His Val Gly Glu Gln Trp

2355 2360 2365

Gln Lys Glu Tyr Leu Gly Ala Ile Cys Ser Cys Thr Cys Phe Gly Gly

2370 2375 2380

Gln Arg Gly Trp Arg Cys Asp Asn Cys Arg Arg Pro Gly Gly Glu Pro

2385 2390 2395 2400

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

2405 2410 2415

Tyr His Gln Arg Thr Asn Thr Asn Val Asn Cys Pro Ile Glu Cys Phe

2420 2425 2430

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

2435 2440 2445

Claims

1.检测与遗传性肾小球疾病相关的突变基因FN1的试剂在制备纤维连接蛋白肾小球病检测试剂盒中的应用，其特征在于，突变基因FN1的核苷酸序列为SEQ ID NO:3，与野生型FN1基因编码DNA的参考序列相比，在基因组位置chr2:216273077处，碱基A突变为碱基G；参考基因组版本是GRCh37。

2.根据权利要求1所述的应用，其特征在于，所述纤维连接蛋白肾小球病检测试剂盒包括引物SEQ ID NO:1和SEQ ID NO:2。

3.根据权利要求2所述的应用，其特征在于，所述纤维连接蛋白肾小球病检测试剂盒还包括Taq DNA聚合酶和PCR缓冲液。