中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2012年
5期
542-546
,共5页
高素青%邓志辉%徐筠娉%王大明%何柳媚%金士正
高素青%鄧誌輝%徐筠娉%王大明%何柳媚%金士正
고소청%산지휘%서균빙%왕대명%하류미%금사정
人类主要组织相容性复合体Ⅰ类相关基因A%双向测序%多态性%基因分型
人類主要組織相容性複閤體Ⅰ類相關基因A%雙嚮測序%多態性%基因分型
인류주요조직상용성복합체Ⅰ류상관기인A%쌍향측서%다태성%기인분형
Human major histocompatibility complex class Ⅰ chain-related gene A%Bi-directional sequencing%Polymorphism%Genotyping
目的 建立稳定的、大规模的MICA基因第2~4外显子双向测序分型检测技术平台,并分析其单核苷酸多态性(single nucleotide polymorphism,SNP).方法 自行设计MICA基因第2~5外显子扩增引物及测序引物,探索PCR扩增及测序的最佳反应条件.用商品化的MICA基因单向测序分型试剂盒作为平行对照,对4个包含MICA* 010等位基因的样本采用自行设计引物扩增,并进行分子克隆和单倍体测序.结果 采用自行建立的MICA基因双向测序分型方法验证了100人份平行对照组单向测序分型结果.应用本研究建立的方法获得了中国人群MICA基因第2~4外显子22个SNP位点.两个新等位基因MICA* 065、MICA* 066获得了世界卫生组织的正式命名.首次发现了MICA等位基因第3内含子新的SNP位点,序列已提交IMGT/H LA数据库.结论 建立的MICA基因双向测序方法可大规模应用于中国人群的MICA基因多态性、组织器官移植配型和疾病研究.
目的 建立穩定的、大規模的MICA基因第2~4外顯子雙嚮測序分型檢測技術平檯,併分析其單覈苷痠多態性(single nucleotide polymorphism,SNP).方法 自行設計MICA基因第2~5外顯子擴增引物及測序引物,探索PCR擴增及測序的最佳反應條件.用商品化的MICA基因單嚮測序分型試劑盒作為平行對照,對4箇包含MICA* 010等位基因的樣本採用自行設計引物擴增,併進行分子剋隆和單倍體測序.結果 採用自行建立的MICA基因雙嚮測序分型方法驗證瞭100人份平行對照組單嚮測序分型結果.應用本研究建立的方法穫得瞭中國人群MICA基因第2~4外顯子22箇SNP位點.兩箇新等位基因MICA* 065、MICA* 066穫得瞭世界衛生組織的正式命名.首次髮現瞭MICA等位基因第3內含子新的SNP位點,序列已提交IMGT/H LA數據庫.結論 建立的MICA基因雙嚮測序方法可大規模應用于中國人群的MICA基因多態性、組織器官移植配型和疾病研究.
목적 건립은정적、대규모적MICA기인제2~4외현자쌍향측서분형검측기술평태,병분석기단핵감산다태성(single nucleotide polymorphism,SNP).방법 자행설계MICA기인제2~5외현자확증인물급측서인물,탐색PCR확증급측서적최가반응조건.용상품화적MICA기인단향측서분형시제합작위평행대조,대4개포함MICA* 010등위기인적양본채용자행설계인물확증,병진행분자극륭화단배체측서.결과 채용자행건립적MICA기인쌍향측서분형방법험증료100인빈평행대조조단향측서분형결과.응용본연구건립적방법획득료중국인군MICA기인제2~4외현자22개SNP위점.량개신등위기인MICA* 065、MICA* 066획득료세계위생조직적정식명명.수차발현료MICA등위기인제3내함자신적SNP위점,서렬이제교IMGT/H LA수거고.결론 건립적MICA기인쌍향측서방법가대규모응용우중국인군적MICA기인다태성、조직기관이식배형화질병연구.
Objective To establish a stable and large-scale bi-directional sequencing platform for genotyping MICA gene exons 2 to 4,and to analyze single nucleotide polymorphisms(SNP) of the region.Methods Primers for particular alleles of MICA gene exons 2 to 5 were designed.Optimal conditions for PCR amplification and sequencing reaction were explored.A commercialized one-way sequencing kit for MICA allele was used as a parallel control.Four samples carrying a MICA * 010 allele were subjected to cloning and haplotype sequencing.Results Results of MICA allele typing of 100 samples for a parallel control group were confirmed by the establish method.Twenty-two SNP in MICA gene exons 2 to 4 were detected in Chinese population.Two novel allelic sequences were accepted by GenBank and IMGT/HLA database and officially named as MICA * 065 and MICA * 066 by the WHO Nomenclature Committee.A novel SNP in MICA gene intron 3 was discovered,with allelic sequence submitted to GenBank and IMGT/HLA database.Conclusion The bi-directional sequencing genotyping platform may be applied for largescale study of MICA allelic polymorphisms,tissue typing,organ transplantation and disease research.