新的HLA-B* 4061等位基因的核苷酸序列分析  

Sequence Analysis of HLA-B* 4061 Allele Newly Found

作　　者：章伟[1,2] 吕沁风[2] 王炜[2] 韩浙东[2] 朱发明[2] 严力行[2] 

机构地区：[1]浙江大学医学院免疫学研究所 [2]浙江省血液中心,卫生部血液安全研究重点实验室,杭州310006 [3]浙江省血液中心,卫生部血液安全研究重点实验室

出　　处：《中国实验血液学杂志》2006年第6期1188-1190,共3页Journal of Experimental Hematology

基　　金：浙江省医药卫生科学研究基金;编号2003Z003

摘　　要：本研究探讨HLA新的等位基因HLA-B* 4061的分子基础。采用盐析法抽提样本DNA,利用PCR方法扩增先证者HLA-B基因的第1-8外显子,PCR产物直接经TOPO克隆到质粒载体中得到单链,对所得克隆进行HLA-B基因第2、3、4外显子双向测序分析。应用PCR-SSP方法证实测序所发现的突变。结果表明先证者样本克隆测序得到2个等位基因,其中1个等位基因为B* 4601,另1个经blast验证其为新的等位基因,新的等位基因序列已递交GenBank(DQ089628,DQ089629,DQ089630)。与最接近的B* 400101等位基因序列相比,新的等位基因仅在第2外显子上有1个核苷酸不同,即第272位C→A,导致第67位氨基酸Ser→Tyr。结论HLA-B* 4061等位基因为新的HLA-B等位基因,已荣获世界卫生组织HLA因子命名委员会正式命名。The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B ＊ 4061, in Chinese population, DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing. The sequencing results showed HLA-B alleles of the proband as B ＊ 4601 and the novel allele. The sequences of the novel allele have been submitted to GenBank （ DQ089628, DQ089629, DQ089630）. After HLA blast analysis, the novel allele showed a single nucleotide mismatch with B ＊ 400101 in exon 2 at position 272 C→A, as the results, changing amino acid from Set to Tyr at codon 67. It is concluded that this allele is a novel one and has been officially named B ＊ 4061 by the WHO Nomenclature Committee.

关 键 词：HLA-B HLA-B＊4061 等位基因 序列分析 

分 类 号：R457.11[医药卫生—治疗学] R457.7[医药卫生—临床医学]