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目的研究南方汉族人群HLA-E基因的分子遗传多态性。方法随机选择150人份南方汉族健康无关个体,采用1对PCR引物特异性扩增HLA-E基因第1-5外显子目的序列,纯化后的PCR扩增产物作为测序模板,对HLA-E基因的第2、3、4外显子进行双向测序,测序反应产物纯化后经ABI3730测序仪电泳,用Assign 3.5 SBT软件分析结果。结果所检测的150例标本均扩增出清晰的目的条带,对HLA-E基因外显子2-4进行双向测序,所获得的序列无背景信号和杂峰。检出了2种HLA-E等位基因,其中HLA-E*01∶01等位基因频率为0.42,HLA-E*01∶03等位基因频率为0.58。3种HLA-E基因型频率的分布符合Hardy-weinberg平衡定律。结论本文所获得了南方汉族人群HLA-E基因分子遗传多态性等基础数据,可为HLA-E的疾病关联研究、临床移植等提供重要参考。
Objective To study the molecular genetic polymorphism of HLA-E gene in southern Chinese Han population. METHODS: A total of 150 unrelated healthy Han Chinese individuals were randomly selected. One pair of PCR primers was used to amplify the specific sequence of exon 1-5 of HLA-E gene. The purified PCR products were used as the sequencing template. HLA-E Genes 2, 3, and 4 exon bi-directional sequencing, sequencing reaction product was purified by ABI3730 sequencer electrophoresis, Assign 3.5 SBT software analysis results. Results All the 150 samples were amplified with a clear target band. The exon 2-4 of HLA-E gene was sequenced bidirectionally, and the obtained sequence had no background signal and no peak. Two kinds of HLA-E alleles were detected, of which HLA-E * 01:01 allele frequency was 0.42, HLA-E * 01:03 allele frequency was 0.58.3 and HLA-E genotype frequencies Distribution in line with Hardy-weinberg equilibrium law. Conclusions The basic data of molecular genetic polymorphism of HLA-E gene in southern Han population obtained in this paper can provide important reference for disease association research and clinical transplantation of HLA-E.