遗传 ›› 2010, Vol. 32 ›› Issue (7): 685-693.doi: 10.3724/SP.J.1005.2010.00685

• 研究报告 • 上一篇    下一篇

中国汉族个体HLA-A、-B基因全长序列的测定及调控区多态性

徐筠娉, 邓志辉, 邹红岩, 高素青, 王大明, 何柳媚, 魏天莉   

  1. 深圳市血液中心, 深圳市组织配型与免疫遗传重点实验室, 深圳 518035
  • 收稿日期:2010-03-01 修回日期:2010-04-27 出版日期:2010-07-20 发布日期:2010-07-20
  • 通讯作者: 徐筠娉 E-mail:yunpingxu1982@yahoo.com.cn
  • 基金资助:

    广东省自然科学基金资助项目(编号:9451803501004124)和广东省科技计划项目(编号:2008B030301277)资助

Cloning and sequencing HLA-A and -B genomic DNA and analyzing polymorphism in regulatory regions in Chinese Han individuals

XU Yun-Ping, DENG Zhi-Hui, ZOU Hong-Yan, GAO Su-Qing, WANG Da-Ming, HE Liu-Mei, WEI Tian-Li   

  1. Shenzhen Key Laboratory of Histocompatibility and Immunogenetics, ShenZhen Blood Center, Shenzhen 518035, China
  • Received:2010-03-01 Revised:2010-04-27 Online:2010-07-20 Published:2010-07-20
  • Contact: XU Yun-Ping E-mail:yunpingxu1982@yahoo.com.cn

摘要: 文章利用20个中国汉族个体样本建立了稳定精确的HLA-A、-B基因全长序列的克隆测序方法, 获得HLA-A 10个等位基因4.2 kb序列, HLA-B 6个等位基因3.7 kb序列, 序列涵盖了两个基因的所有外显子、所有内含子、5′启动子区以及3′非翻译区(3′UTR)。A*1153是文章发现的一个新等位基因, B*151101的内含子序列、5个HLA-A以及2个HLA-B等位基因的5′启动子序列和3′UTR序列为国际上首次报道, 其他等位基因均延伸了IMGT/HLA数据库中释放的全长序列。文章首次在中国汉族个体中测定了IMGT/HLA数据库中没有覆盖的HLA-A、-B基因的上游5′启动子以及下游3′UTR区域的多态性模式。HLA-A基因5′启动子延伸区域共发现26个SNPs和一处3 bp(AAA/-)的插入/缺失, 3′UTR延伸区域共发现14个SNPs; HLA-B基因5′启动子延伸区域共发现5个SNPs和一处1 bp(T/-)的插入/缺失, 3′UTR延伸区域共发现8个SNPs。通过对两个基因的5′启动子、外显子以及3′UTR的系统发育树分析, 发现两个基因调控区与外显子的进化关系有所不同, HLA-A基因除A*24020101外, 其他等位基因两端调控区与外显子连锁比较紧密, HLA-B基因两端调控区与外显子之间则发生了较为频繁的重组事件。

关键词: 人类白细胞抗原, 基因组序列, 克隆测序, 调控区, 多态性

Abstract: In the present study, a high-resolute method for cloning and sequencing genomic full-length HLA-A and -B using 20 Chinese Han individuals was established. We detected 10 HLA-A allele sequences 4.2 kb in length and 6 HLA-B allele sequences 3.7 kb in length, and the sequences included all exons, all introns, 5′promoter, and 3′UTR of the two genes. All sixteen sequences have been submitted to GenBank and IMGT/HLA database. A*1153 is a novel allele, and the introns of B*151101 are firstly reported here. The 5′promoter and 3′UTR sequences of 5 HLA-A alleles and 2 HLA-B alleles are also firstly disclosed, and all other alleles have extended the genomic full length sequences released in IMGT/HLA database. The polymorphic structures of upper 5′promoter and downstream 3′UTR, which were uncovered in IMGT/HLA database, are firstly depicted in Chinese Han individuals. Twenty-six single nucleotide polymorphisms (SNPs) and one 3 bp-insertion/deletion (Indel) were located in the upper 5′promoter and 14 SNPs were located in the 3′UTR of HLA-A. In addition, five SNPs and one 1 bp-indel were located in the upper 5′promoter and 5 SNPs were located in the 3′UTR of HLA-B. Through analyzing the phylogenetic trees of 5′promoter, exons and 3′UTR of the two genes, we found that the evolution history of regulatory regions and exons is different between the two genes. The regulatory regions are tightly linked with exons in most of HLA-A alleles excluding A*24020101. On the contrary, recombinant events may occur fre-quently between regulatory regions and exons in most HLA-B alleles.

Key words: polymorphism, human leucocyte antigen (HLA), genomic sequence, cloning and sequencing, regulatory region