一例新的HLA-B等位基因B*5614的核苷酸序列分析  

Identification and sequence analysis of a novel HLA-B*5614 a llele

作　　者：朱发明[1] 吕沁风[1] 章伟[1] 张海琴[1] 傅启华[1] 严力行[1] 

机构地区：[1]浙江省血液中心,杭州310006

出　　处：《中华医学遗传学杂志》2005年第3期288-290,共3页Chinese Journal of Medical Genetics

基　　金：浙江省医药卫生科学研究基金(2003Z003)~~

摘　　要：目的研究HLA新的等位基因HLAB5614的分子基础。方法样本DNA抽提采用盐析法,利用PCR方法扩增先证者HLAB基因的第2～4外显子,PCR产物直接经TOPO转染克隆到质粒载体中分离其等位基因,对所得克隆进行第2～4外显子双向测序分析。应用序列特异性引物PCR方法证实测序所发现的突变。结果先证者样本克隆测序得到两个等位基因,其中1个等位基因为B1502,另一个经BLAST验证为新的等位基因,新的等位基因序列已递交GenBank(AY601726,AY601727,AY601728)。与最接近的B5608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277位G→C,导致第93位氨基酸Gly→Arg。结论该等位基因为新的HLAB等位基因,被世界卫生组织HLA因子命名委员会正式命名为HLAB5614。Objective To investigate the molecular genetics b asis for a novel HLA allele, HLA-B*5614, in Chinese population. Methods DNA was extracted from whole blood by salting-out method. The HLA-B exon s 2~4 of the proband was amplified and the amplified product was cloned using TO PO 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 conf irm the mutations detected by sequencing. Results The sequencin g results showed the HLA-B alleles of the proband as B*1502 and the novel allel e. T he sequences of the novel allele have been submitted to GenBank(AY601726,AY610 727,AY610728).After BLAST analysis, the novel allele differs from B*5608 by a single nucleotide at position 277G→C in exon 2. This results in an amino acid c hange from Gly to Arg at codon 93. Conclusion This allele is a novel allele and has been officially named B*5614 by the WHO Nomenclature Commi ttee.

关 键 词：HLA-B 等位基因 B＊5614 核苷酸序列 遗传因素 

分 类 号：R394[医药卫生—医学遗传学]