CN108342471B - 常染色体显性非综合征听力损失耳聋osbpl2基因突变体 - Google Patents

常染色体显性非综合征听力损失耳聋osbpl2基因突变体 Download PDF

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CN108342471B
CN108342471B CN201810260317.7A CN201810260317A CN108342471B CN 108342471 B CN108342471 B CN 108342471B CN 201810260317 A CN201810260317 A CN 201810260317A CN 108342471 B CN108342471 B CN 108342471B
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吴柒柱
白海花
杨立清
李星
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Abstract

本发明涉及生物领域，特别涉及一种常染色体显性非综合征听力损失耳聋OSBPL2基因突变体。耳聋病因主要有遗传因素、环境因素和其他不明因素，遗传方式主要包括常染色体显性遗传、常染色体隐性遗传和X‑连锁等，均属单基因遗传病。本发明提供了一种常染色体显性非综合征听力损失耳聋OSBPL2基因突变体新的突变位点：c.158_159delAA；p.Gln53Arg fs*100，为杂合子，推测OSBPL2基因为常染色体显性非综合征听力损失耳聋的致病基因。该突变基因可应用于婚前遗传咨询，基于DNA的产前诊断以及症状前诊断，制备非综合征型听力损失耳聋检测试剂等方面，进一步为该类疾病的早期筛查和诊断提供依据，可减少降低发病率，达到提高人口素质的目的。

Description

常染色体显性非综合征听力损失耳聋OSBPL2基因突变体
技术领域
本发明涉及生物领域中的疾病相关突变基因，特别涉及一遗传性迟发型耳聋大家系常染色体显性非综合征型耳聋相关的OSBPL2基因的新的突变位点。
背景技术
耳聋是人类感觉系统缺陷，严重影响人类健康。耳聋病因主要有遗传因素、环境因素和其他不明因素。其中约50%~60%的耳聋患者为遗传性耳聋。遗传方式主要包括常染色体显性遗传、常染色体隐性遗传和X-连锁等，均属单基因遗传病。目前，已发现常染色体显性遗传性耳聋（autosomal dominantnon-syndromic sensorineural hearing loss, DFNA）基因位点67个，已克隆34个（http://hereditaryhearingloss.org.2017年），尚有一半有待克隆。因此，本领域仍然需要发现DFNA相关的基因突变，以及检测相关突变基因的引物、试剂盒和方法。
伴随人类基因组计划的完成，人类基因组图谱已基本构建完成。本研究采用二代测序技术，对先证者及家系中另外两名患者及两名正常个体进行全基因组测序，获得候选致病基因，针对该突变基因，对家系患者及正常个体进行Sanger 测序验证并确认。
发明内容
本发明所要解决的技术问题是提供一种常染色体显性非综合征听力损失（ADNSHL）耳聋相关的OSBPL2基因突变体。
本发明所提供的常染色体显性非综合征听力损失耳聋OSBPL2基因突变体新的突变位点是：在SEQ ID NO:1 OSBPL2基因编码区第158和159位发生两个腺嘌呤核苷酸缺失（c.158_159delAA；p.Gln53Arg fs*100），为杂合子。序列分析表明该位置缺失突变引起OSBPL2基因编码蛋白发生移码突变，移码阅读框的第100位密码子突变成为终止密码子（UAA），导致原野生型SEQ ID NO:2 OSBPL2蛋白中99个氨基酸残基53-151位改变，329个氨基酸残基缺失，突变为SEQ ID NO:5氨基酸序列。据报道，OSBPL2 蛋白的生物学功能为主要参与细胞胆固醇转运与脂质代谢等生理功能，经过SIFT和PolyPhen等蛋白功能预测显示，大量氨基酸的改变和缺失将影响其蛋白功能。没有发现该位点致病性的报道及在人群中的频率。推测OSBPL2基因为常染色体显性非综合征听力损失耳聋的致病基因。
本发明提供的与非综合征型遗传性耳聋的新的突变基因可应用于婚前遗传咨询，基于DNA 的产前诊断以及症状前诊断，制备非综合征型听力损失耳聋检测试剂等方面，可减少降低发病率，达到提高人口素质的目的。
本发明的有益效果是：对一个遗传性迟发型耳聋大家系进行临床表型及家系遗传特征分析，并对常见耳聋基因及位点进行初步筛查，进一步为该类疾病的早期筛查和诊断提供依据。
附图说明
本申请附图包括：
图1、一遗传性迟发型耳聋大家系ADNSHL耳聋的系谱图；
（↗为先证者、■男性受累者、●女性受累者、□男性表型正常者、○女性表型正常者；
*为高通量测序对象：Ⅳ-2、Ⅳ-9、Ⅳ-14、Ⅴ-6、Ⅴ-9）
图2、先证者Ⅴ-9与对照Ⅴ-6纯音听力检测图；
（A和B为患者Ⅴ-9，听力损伤；C和D为Ⅴ-6，正常听力）
图3、OSBPL2基因结构图；
图4、先证者Ⅴ-9及患者Ⅳ-2、Ⅳ-9、Ⅳ-12、Ⅴ-7、Ⅴ-11、Ⅵ-5测序图；
（图中竖线处为突变位置）
图5、家系正常成员Ⅳ-14、Ⅴ-6、Ⅴ-13、Ⅵ-1、Ⅵ-4、Ⅵ-6测序图；
（图中竖线处为家系正常成员与患者突变的位置比较）。
具体实施方式
1、样本来源
研究样本来自位于内蒙古西部地区，已传6代的大家系，共计53人，其中耳聋患者19例： 12例男性患者， 7例女性患者。已绘制该家系系谱图，如图1。所有受调查者均签署知情同意书，符合内蒙古民族大学附属医院伦理委员会伦理论证要求。该病在这一家系中已传6代，受试者包括先证者在内共计13例，其中耳聋患者为7例，听力正常者为6例，进行病史采集、听力学检测、全身体格检查和生活习惯调查等，临床诊断发现，该家系耳聋患者临床表现均为迟发性听力下降，双侧对称性轻度至重度感音神经性耳聋。初步诊断为常染色体显性非综合征听力损失耳聋。图1中，6代非综合征型遗传性耳聋系谱中：↗为先证者；■男性受累者；●女性受累者；□男性表型正常者；○女性表型正常者。
2、研究样本的特征
研究样本为内蒙古西部地区一大家系，6代患有非综合征型遗传性耳聋家系，表现为常染色体显性非综合征听力损失(ADNSHL)，现生存受累患者14例。该样本中的受累成员（18~62岁）临床表现均为迟发性听力下降，发病年龄在10~40岁，均有耳鸣病史，无前庭功能障碍，无耳毒性药物接触史，无噪声接触史，均无智力障碍。听力损失均表现为语后聋，听力损失程度随年龄而加剧。听力检测诊断为双侧对称性轻度至重度感音神经性耳聋，听力曲线多为平坦型，如图2。该家系每一代都有患者而且男女均发病，属于常染色体显性方式遗传。
3、突变基因及其检测方法的获得
采用诺禾致源Illumina平台进行样本（Ⅳ-2;Ⅳ-9;Ⅳ-14;Ⅴ-6;Ⅴ-9）全基因组测序分析，结果如下：
先证者（Ⅴ-9）OSBPL2基因编码区158和159位发生两个腺嘌呤核苷酸缺失（c.158_159delAA；p.Gln53Arg fs*100），为杂合子。序列分析表明该位置缺失突变引起OSBPL2基因编码蛋白发生移码突变，移码阅读框的第100位密码子成为终止密码子（UAA），导致原野生型OSBPL2蛋白中99个氨基酸残基（53-151位）改变，329个氨基酸残基缺失。经过SIFT和PolyPhen等蛋白功能预测显示，大量氨基酸的改变和缺失将影响其蛋白功能。没有发现该位点致病性的报道及在人群中的频率。推测OSBPL2基因该突变位点为ADNSHL耳聋的致病基因的又一突变类型。
野生型OSBPL2基因的cDNA序列如SEQ ID NO:1所示。野生型OSBPL2蛋白的氨基酸序列如SEQ ID NO:2所示。突变型OSBPL2蛋白的氨基酸序列如SEQ ID NO:5所示。
另外，OSBPL2基因的其他3个突变：c.141_142delTG(p.Arg50Ala fs*103)、c.153_154delCT(p.Gln53Arg fs*100)、c.583C>A(p.Leu195Met)，在本申请高通量测序对象中均未发现。
4、突变基因PCR验证
应用聚合酶链反应产物直接测序方法，对包括先证者在内的13名家系成员的OSBPL2基因编码区的Chr20:60835157-60835158（或OSBPL2基因cDNA的158和159位）c.158_159delAA（OSBPL2基因 cDNA158的A和159位的A缺失）进行突变检测，引物序列如下：
SEQ ID NO:3——Sense primer（上游）：5' -TCAGGTCCAGCGAAAATG -3'
SEQ ID NO:4——Anti-sense primer（下游）：5' -TTAGATGGGGAAAGGCAC -3'
PCR 产物测序结果与 GenBank 登录OSBPL2（NG_042164.1）基因序列比对，结果显示表型正常者OSBPL2基因序列与 GenBank给定序列完全一致。而非综合征型遗传性耳聋患者与家系中正常表型者比较，患者OSBPL2基因编码区存在 1 个杂合移码突变c.158_159delAA，存在家系共分离。由此判断，此蒙古族常显性非综合征型遗传性耳聋的致病突变位点为OSBPL2基因上的c.158_159delAA突变位点。
通过全外显子测序数据分析确定1个潜在基因致病突变位点，后续经直接测序验证，此突变位点在家系中存在共分离现象。13名受试者测序结果为：7例耳聋患者发生此位点突变；6名听力正常者均没有此位点突变。
5、细胞和分子功能实验
利用OSBPL2基因（野生型和突变型）在Huh7 细胞内定位并过表达对脂类，尤其是胆固醇代谢的影响以及与DIAPH1蛋白的相互作用，证实了OSBPL2基因为导致ADNSHL耳聋的新致病基因，该位点突变后可能通过影响毛细胞的转导功能从而引起听力的丧失，但具体机制有待于进一步的深入研究。
【序列表说明】
以下对上述核苷酸和氨基酸序列进行说明：
SEQ ID NO:1——野生型OSBPL2基因的cDNA序列
ATGAACGGAG AGGAAGAATT CTTTGATGCC GTCACAGGCT TTGATTCTGA TAACTCTTCT60
GGGGAATTTT CAGAGGCAAA TCAGAAAGTC ACGGGAATGA TTGACTTAGA CACCAGCAAA120
AATAATAGGA TTGGGAAAAC TGGGGAGAGG CCCTCTCAAG AGAACGGAAT TCAGAAACAC180
AGGACATCGC TGCCGGCTCC CATGTTCAGC AGAAGCGACT TCAGCGTGTG GACCATCCTG240
AAGAAGTGTG TTGGCCTGGA GCTGTCCAAG ATCACGATGC CAATCGCCTT CAACGAGCCT300
CTGAGCTTCT TGCAGCGGAT CACGGAGTAC ATGGAGCACG TGTACCTCAT CCACAGGGCC 360
TCCTGCCAGC CCCAGCCCCT GGAGAGGATG CAGTCTGTGG CTGCTTTTGC TGTTTCGGCT 420
GTGGCTTCCC AGTGGGAGAG GACCGGCAAA CCATTTAATC CACTCTTGGG AGAAACGTAT 480
GAATTAATCA GGGAAGATTT AGGATTCAGA TTTATATCGG AACAGGTCAG TCACCACCCC 540
CCCATCAGTG CGTTCCACTC GGAAGGTCTC AACCATGACT TCCTGTTCCA TGGCTCCATC 600
TACCCCAAGC TCAAGTTCTG GGGCAAAAGC GTGGAGGCGG AGCCCCGAGG CACCATCACC 660
CTGGAGCTGC TCAAACATAA TGAAGCCTAC ACCTGGACCA ACCCCACCTG CTGCGTCCAC 720
AACGTCATCA TCGGGAAGCT GTGGATAGAG CAGTATGGGA CAGTGGAGAT TTTAAACCAC 780
AGAACTGGAC ATAAGTGTGT GCTTCACTTT AAACCGTGTG GATTATTTGG AAAAGAACTT 840
CACAAGGTGG AAGGACACAT TCAAGACAAA AACAAAAAGA AGCTCTTTAT GATCTATGGC 900
AAATGGACGG AATGTTTGTG GGGCATAGAT CCTGTTTCGT ATGAATCCTT CAAGAAGCAG 960
GAGAGGAGAG GTGACCACCT GAGAAAGGCC AAGCTGGATG AAGACTCCGG GAAGGCTGAC 1020
AGCGACGTGG CTGACGACGT GCCTGTGGCC CAGGAGACCG TGCAGGTCAT TCCTGGCAGC 1080
AAGCTGCTCT GGAGGATCAA CACCCGGCCC CCCAACTCTG CCCAGATGTA TAATTTCACC 1140
AGTTTCACTG TGAGCCTCAA CGAGCTGGAG ACAGGCATGG AGAAGACCCT GCCACCCACG 1200
GACTGCCGCC TGCGCCCTGA CATCCGCGGC ATGGAGAATG GCAACATGGA TCTGGCCAGC 1260
CAGGAGAAGG AGCGGCTGGA GGAGAAGCAG AGAGAAGCAC GGAGGGAGCG GGCCAAGGAG 1320
GAGGCAGAGT GGCAGACGAG GTGGTTCTAC CCAGGCAATA ACCCCTACAC TGGGACCCCC 1380
GACTGGTTGT ATGCAGGGGA TTACTTTGAG CGGAATTTCT CCGACTGCCC AGATATCTAT 1440
TAA 1443
SEQ ID NO:2——野生型OSBPL2蛋白的氨基酸序列
Met Asn Gly Glu Glu Glu Phe Phe Asp Ala Val Thr Gly Phe Asp Ser
1 5 10 15
Asp Asn Ser Ser Gly Glu Phe Ser Glu Ala Asn Gln Lys Val Thr Gly
20 25 30
Met Ile Asp Leu Asp Thr Ser Lys Asn Asn Arg Ile Gly Lys Thr Gly
35 40 45
Glu Arg Pro Ser Gln Glu Asn Gly Ile Gln Lys His Arg Thr Ser Leu
50 55 60
Pro Ala Pro Met Phe Ser Arg Ser Asp Phe Ser Val Trp Thr Ile Leu
65 70 75 80
Lys Lys Cys Val Gly Leu Glu Leu Ser Lys Ile Thr Met Pro Ile Ala
85 90 95
Phe Asn Glu Pro Leu Ser Phe Leu Gln Arg Ile Thr Glu Tyr Met Glu
100 105 110
His Val Tyr Leu Ile His Arg Ala Ser Cys Gln Pro Gln Pro Leu Glu
115 120 125
Arg Met Gln Ser Val Ala Ala Phe Ala Val Ser Ala Val Ala Ser Gln
130 135 140
Trp Glu Arg Thr Gly Lys Pro Phe Asn Pro Leu Leu Gly Glu Thr Tyr
145 150 155 160
Glu Leu Ile Arg Glu Asp Leu Gly Phe Arg Phe Ile Ser Glu Gln Val
165 170 175
Ser His His Pro Pro Ile Ser Ala Phe His Ser Glu Gly Leu Asn His
180 185 190
Asp Phe Leu Phe His Gly Ser Ile Tyr Pro Lys Leu Lys Phe Trp Gly
195 200 205
Lys Ser Val Glu Ala Glu Pro Arg Gly Thr Ile Thr Leu Glu Leu Leu
210 215 220
Lys His Asn Glu Ala Tyr Thr Trp Thr Asn Pro Thr Cys Cys Val His
225 230 235 240
Asn Val Ile Ile Gly Lys Leu Trp Ile Glu Gln Tyr Gly Thr Val Glu
245 250 255
Ile Leu Asn His Arg Thr Gly His Lys Cys Val Leu His Phe Lys Pro
260 265 270
Cys Gly Leu Phe Gly Lys Glu Leu His Lys Val Glu Gly His Ile Gln
275 280 285
Asp Lys Asn Lys Lys Lys Leu Phe Met Ile Tyr Gly Lys Trp Thr Glu
290 295 300
Cys Leu Trp Gly Ile Asp Pro Val Ser Tyr Glu Ser Phe Lys Lys Gln
305 310 315 320
Glu Arg Arg Gly Asp His Leu Arg Lys Ala Lys Leu Asp Glu Asp Ser
325 330 335
Gly Lys Ala Asp Ser Asp Val Ala Asp Asp Val Pro Val Ala Gln Glu
340 345 350
Thr Val Gln Val Ile Pro Gly Ser Lys Leu Leu Trp Arg Ile Asn Thr
355 360 365
Arg Pro Pro Asn Ser Ala Gln Met Tyr Asn Phe Thr Ser Phe Thr Val
370 375 380
Ser Leu Asn Glu Leu Glu Thr Gly Met Glu Lys Thr Leu Pro Pro Thr
385 390 395 400
Asp Cys Arg Leu Arg Pro Asp Ile Arg Gly Met Glu Asn Gly Asn Met
405 410 415
Asp Leu Ala Ser Gln Glu Lys Glu Arg Leu Glu Glu Lys Gln Arg Glu
420 425 430
Ala Arg Arg Glu Arg Ala Lys Glu Glu Ala Glu Trp Gln Thr Arg Trp
435 440 445
Phe Tyr Pro Gly Asn Asn Pro Tyr Thr Gly Thr Pro Asp Trp Leu Tyr
450 455 460
Ala Gly Asp Tyr Phe Glu Arg Asn Phe Ser Asp Cys Pro Asp Ile Tyr
465 470 475 480
SEQ ID NO:3——Sense primer（上游）：5' -TCAGGTCCAGCGAAAATG -3'
SEQ ID NO:4——Anti-sense primer（下游）：5' -TTAGATGGGGAAAGGCAC -3'
SEQ ID NO:5——突变型OSBPL2蛋白的氨基酸序列
Met Asn Gly Glu Glu Glu Phe Phe Asp Ala Val Thr Gly Phe Asp Ser
1 5 10 15
Asp Asn Ser Ser Gly Glu Phe Ser Glu Ala Asn Gln Lys Val Thr Gly
20 25 30
Met Ile Asp Leu Asp Thr Ser Lys Asn Asn Arg Ile Gly Lys Thr Gly
35 40 45
Glu Arg Pro Ser Arg Glu Arg Asn Ser Glu Thr Gln Asp Ile Ala Ala
50 55 60
Gly Ser His Val Gln Gln Lys Arg Leu Gln Arg Val Asp His Pro Glu
65 70 75 80
Glu Val Cys Trp Pro Gly Ala Val Gln Asp His Asp Ala Asn Arg Leu
85 90 95
Gln Arg Ala Ser Glu Leu Leu Ala Ala Asp His Gly Val His Gly Ala
100 105 110
Arg Val Pro His Pro Gln Gly Leu Leu Pro Ala Pro Ala Pro Gly Glu
115 120 125
Asp Ala Val Cys Gly Cys Phe Cys Cys Phe Gly Cys Gly Phe Pro Val
130 135 140
Gly Glu Asp Arg Gln Thr Ile
145 150
序列表
<110> 吴柒柱
<120> 常染色体显性非综合征听力损失耳聋OSBPL2基因突变体
<160> 5
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1443
<212> DNA
<213> Human
<400> 1
atgaacggag aggaagaatt ctttgatgcc gtcacaggct ttgattctga taactcttct 60
ggggaatttt cagaggcaaa tcagaaagtc acgggaatga ttgacttaga caccagcaaa 120
aataatagga ttgggaaaac tggggagagg ccctctcaag agaacggaat tcagaaacac 180
aggacatcgc tgccggctcc catgttcagc agaagcgact tcagcgtgtg gaccatcctg 240
aagaagtgtg ttggcctgga gctgtccaag atcacgatgc caatcgcctt caacgagcct 300
ctgagcttct tgcagcggat cacggagtac atggagcacg tgtacctcat ccacagggcc 360
tcctgccagc cccagcccct ggagaggatg cagtctgtgg ctgcttttgc tgtttcggct 420
gtggcttccc agtgggagag gaccggcaaa ccatttaatc cactcttggg agaaacgtat 480
gaattaatca gggaagattt aggattcaga tttatatcgg aacaggtcag tcaccacccc 540
cccatcagtg cgttccactc ggaaggtctc aaccatgact tcctgttcca tggctccatc 600
taccccaagc tcaagttctg gggcaaaagc gtggaggcgg agccccgagg caccatcacc 660
ctggagctgc tcaaacataa tgaagcctac acctggacca accccacctg ctgcgtccac 720
aacgtcatca tcgggaagct gtggatagag cagtatggga cagtggagat tttaaaccac 780
agaactggac ataagtgtgt gcttcacttt aaaccgtgtg gattatttgg aaaagaactt 840
cacaaggtgg aaggacacat tcaagacaaa aacaaaaaga agctctttat gatctatggc 900
aaatggacgg aatgtttgtg gggcatagat cctgtttcgt atgaatcctt caagaagcag 960
gagaggagag gtgaccacct gagaaaggcc aagctggatg aagactccgg gaaggctgac 1020
agcgacgtgg ctgacgacgt gcctgtggcc caggagaccg tgcaggtcat tcctggcagc 1080
aagctgctct ggaggatcaa cacccggccc cccaactctg cccagatgta taatttcacc 1140
agtttcactg tgagcctcaa cgagctggag acaggcatgg agaagaccct gccacccacg 1200
gactgccgcc tgcgccctga catccgcggc atggagaatg gcaacatgga tctggccagc 1260
caggagaagg agcggctgga ggagaagcag agagaagcac ggagggagcg ggccaaggag 1320
gaggcagagt ggcagacgag gtggttctac ccaggcaata acccctacac tgggaccccc 1380
gactggttgt atgcagggga ttactttgag cggaatttct ccgactgccc agatatctat 1440
taa 1443
<210> 2
<211> 480
<212> PRT
<213> Human
<400> 2
Met Asn Gly Glu Glu Glu Phe Phe Asp Ala Val Thr Gly Phe Asp Ser
1 5 10 15
Asp Asn Ser Ser Gly Glu Phe Ser Glu Ala Asn Gln Lys Val Thr Gly
20 25 30
Met Ile Asp Leu Asp Thr Ser Lys Asn Asn Arg Ile Gly Lys Thr Gly
35 40 45
Glu Arg Pro Ser Gln Glu Asn Gly Ile Gln Lys His Arg Thr Ser Leu
50 55 60
Pro Ala Pro Met Phe Ser Arg Ser Asp Phe Ser Val Trp Thr Ile Leu
65 70 75 80
Lys Lys Cys Val Gly Leu Glu Leu Ser Lys Ile Thr Met Pro Ile Ala
85 90 95
Phe Asn Glu Pro Leu Ser Phe Leu Gln Arg Ile Thr Glu Tyr Met Glu
100 105 110
His Val Tyr Leu Ile His Arg Ala Ser Cys Gln Pro Gln Pro Leu Glu
115 120 125
Arg Met Gln Ser Val Ala Ala Phe Ala Val Ser Ala Val Ala Ser Gln
130 135 140
Trp Glu Arg Thr Gly Lys Pro Phe Asn Pro Leu Leu Gly Glu Thr Tyr
145 150 155 160
Glu Leu Ile Arg Glu Asp Leu Gly Phe Arg Phe Ile Ser Glu Gln Val
165 170 175
Ser His His Pro Pro Ile Ser Ala Phe His Ser Glu Gly Leu Asn His
180 185 190
Asp Phe Leu Phe His Gly Ser Ile Tyr Pro Lys Leu Lys Phe Trp Gly
195 200 205
Lys Ser Val Glu Ala Glu Pro Arg Gly Thr Ile Thr Leu Glu Leu Leu
210 215 220
Lys His Asn Glu Ala Tyr Thr Trp Thr Asn Pro Thr Cys Cys Val His
225 230 235 240
Asn Val Ile Ile Gly Lys Leu Trp Ile Glu Gln Tyr Gly Thr Val Glu
245 250 255
Ile Leu Asn His Arg Thr Gly His Lys Cys Val Leu His Phe Lys Pro
260 265 270
Cys Gly Leu Phe Gly Lys Glu Leu His Lys Val Glu Gly His Ile Gln
275 280 285
Asp Lys Asn Lys Lys Lys Leu Phe Met Ile Tyr Gly Lys Trp Thr Glu
290 295 300
Cys Leu Trp Gly Ile Asp Pro Val Ser Tyr Glu Ser Phe Lys Lys Gln
305 310 315 320
Glu Arg Arg Gly Asp His Leu Arg Lys Ala Lys Leu Asp Glu Asp Ser
325 330 335
Gly Lys Ala Asp Ser Asp Val Ala Asp Asp Val Pro Val Ala Gln Glu
340 345 350
Thr Val Gln Val Ile Pro Gly Ser Lys Leu Leu Trp Arg Ile Asn Thr
355 360 365
Arg Pro Pro Asn Ser Ala Gln Met Tyr Asn Phe Thr Ser Phe Thr Val
370 375 380
Ser Leu Asn Glu Leu Glu Thr Gly Met Glu Lys Thr Leu Pro Pro Thr
385 390 395 400
Asp Cys Arg Leu Arg Pro Asp Ile Arg Gly Met Glu Asn Gly Asn Met
405 410 415
Asp Leu Ala Ser Gln Glu Lys Glu Arg Leu Glu Glu Lys Gln Arg Glu
420 425 430
Ala Arg Arg Glu Arg Ala Lys Glu Glu Ala Glu Trp Gln Thr Arg Trp
435 440 445
Phe Tyr Pro Gly Asn Asn Pro Tyr Thr Gly Thr Pro Asp Trp Leu Tyr
450 455 460
Ala Gly Asp Tyr Phe Glu Arg Asn Phe Ser Asp Cys Pro Asp Ile Tyr
465 470 475 480
<210> 3
<211> 18
<212> DNA
<213> Human
<400> 3
tcaggtccag cgaaaatg 18
<210> 4
<211> 18
<212> DNA
<213> Human
<400> 4
ttagatgggg aaaggcac 18
<210> 5
<211> 151
<212> PRT
<213> Human
<400> 5
Met Asn Gly Glu Glu Glu Phe Phe Asp Ala Val Thr Gly Phe Asp Ser
1 5 10 15
Asp Asn Ser Ser Gly Glu Phe Ser Glu Ala Asn Gln Lys Val Thr Gly
20 25 30
Met Ile Asp Leu Asp Thr Ser Lys Asn Asn Arg Ile Gly Lys Thr Gly
35 40 45
Glu Arg Pro Ser Arg Glu Arg Asn Ser Glu Thr Gln Asp Ile Ala Ala
50 55 60
Gly Ser His Val Gln Gln Lys Arg Leu Gln Arg Val Asp His Pro Glu
65 70 75 80
Glu Val Cys Trp Pro Gly Ala Val Gln Asp His Asp Ala Asn Arg Leu
85 90 95
Gln Arg Ala Ser Glu Leu Leu Ala Ala Asp His Gly Val His Gly Ala
100 105 110
Arg Val Pro His Pro Gln Gly Leu Leu Pro Ala Pro Ala Pro Gly Glu
115 120 125
Asp Ala Val Cys Gly Cys Phe Cys Cys Phe Gly Cys Gly Phe Pro Val
130 135 140
Gly Glu Asp Arg Gln Thr Ile
145 150

Claims (2)

1.检测OSBPL2基因突变体的试剂在制备常染色体显性非综合征听力损失耳聋的检测试剂中的应用，其特征在于：
该突变体的突变位点是：OSBPL2基因编码区第158和159位发生两个腺嘌呤核苷酸缺失，具体为：c.158_159delAA；p.Gln53Arg fs*100，为杂合子，所述OSBPL2基因编码区的核苷酸序列如SEQ ID NO:1所示。
2.根据权利要求1所述的应用，其特征在于：
所述位点发生缺失突变引起OSBPL2基因编码蛋白发生移码突变，移码阅读框的第100位密码子突变成为终止密码子UAA，导致野生型OSBPL2蛋白中99个氨基酸残基53-151位改变，329个氨基酸残基缺失，突变型OSBPL2蛋白的氨基酸序列如SEQ ID NO:5所示。
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