HLA-DRB1＊1218新等位基因克隆测序鉴定及内含子序列分析  

Identification of a novel allele HLA-DRB1 ＊ 1218

作　　者：高素青[1] 邓志辉[1] 徐筠娉[1] 

机构地区：[1]广东省深圳市血液中心研究所,中华骨髓库深圳HLA分型实验室, 518035

出　　处：《中华医学遗传学杂志》2009年第3期272-276,共5页Chinese Journal of Medical Genetics

基　　金：深圳市科技计划项目（200603202）

摘　　要：目的鉴定中国人群人类白细胞抗原（human leukocyte antigen，HLA）DRB1＊1218新等位基因，分析新等位基因第1和2内含子序列信息。方法采用聚合酶链反应一序列特异寡核苷酸探针反向杂交法（polymerase chain reaction-sequence specific oligonucleotide probes，PCR—SSOP）对广东地区随机正常人群进行HLA常规基因分型，发现1个与HI。A-DRB1＊120201相近的未知基因，对先证者DNA应用组特异性引物扩增HLA—DRB1位点第2外显子，PCR产物经克隆到质粒载体中以获得单链，对克隆所得产物进行HLA—DRB1基因的第2外显子及第1和第2内含子双向测序分析，并与DRBl＊120201基因序列的第2外显子和DRBl＊03010101等位基因内含子相比较。结果发现该个体的一个HLA—DRB1＊080302基因被确认，而另一个HI。A—DRB1基因为新等位基因，其序列被GenBank接受（编号为FJ481086）。新等位基因与最相近的DRB1＊120201相比，在第2外显子上有1个核苷酸的不同，即第262位G—C（密码子59GAG—CAG，氨基酸59G1u—GIn）。DRB1＊1218与DRB1＊03010101等位基因第2内含子序列完全相同，而与DRB1＊03010101等位基因第1内含子序列相比较有12个碱基不同。结论发现并鉴定一个新的HLA等位基因，经世界卫生组织HLA因子命名委员会正式命名为HLA—DRB1＊1218．Objective To identify a novel human leukocyte antigen （HLA） allele by cloning and sequence-based typing in Chinese population, and analyzing the sequence of the introns 1 and 2. Methods The routine HLA-A, B, DRB1 low resolution genotyping for stem cell donor from Guangdong province was performed with polymerase chain reaction-sequence specific oligonucleotide probes （PCR-SSOP）. An unknown HLA-DRB1 allele was initially detected by HLA typing. Genomic DNA of the proband was amplified by using HLA-DRB1 locus group-specific primer, the amplified product was cloned, sequenced, and compared to the closest DRB1 ＊ 120201 allele and the closest intron sequence of the DRB1 ＊ 030101 allele. Results The sequencing results showed that a normal DRB1 ＊ 080302 and a novel DRB1 ＊ 1218 variant allele were identified. The sequence of the novel allele has been submitted to GenBank （FJ481086）. The novel allele had 1 nucleotide substitution of the closest matching allele HLA-DRB1 ＊ 120201 at nt262（G→C） in exon 2, resulting in an amino acid change from Glu（GAG）→Gln （CAG） at codon 59. The intron 2 sequence is identical between the novel HLA-DRB1 ＊ 1218 and DRB1 ＊ 030101, but there are 12 nucleotides substitution in intron 1. Conclusion A novel HLA allele was confirmed by cloning and sequence-based typing in Chinese. It was officially designated as HLA-DRB1＊1218 by WHO Nomenclature Committee.

关 键 词：人类白细胞抗原 等位基因 外显子 内含子 

分 类 号：R686[医药卫生—骨科学]