CAJ | 학술논문

目的研究人类白细胞抗原(human leukocyte antigen,HLA)新等位基因HLA_B*9536和B*4612的分子机制.方法采用Invitrogen抽提试剂盒抽提标本DNA,利用单链特异性引物PCR方法扩增样本HLA-B基因第2～4外显子,对PCR产物直接进行HLA-B基因第2、3、4外显子双向测序分析.结果先证者标本存在2个HLA-B等位基因,经HLA Blast验证均为新的等位基因,新的等位基因序列已递交GenBank(EU081878和EU081879),经世界卫生组织HLA命名委员会正式命名为HLA-B*9536和HLA-B*4612.HLA-B*9536第2～4外显子序列与最接近的B*1505相比,在第3外显子存在一个碱基的不同,即第544位G→A改变,导致第158位氨基酸Ala→Thr;HLA-B*4612第2～4外显子序列与最接近的B*4601相比,在第3外显子存在一个碱基的不同,即第363位G→C,导致第97位氨基酸Arg→Ser.结论在同一标本中发现两个新的HLA-B等位基因,并被世界卫生组织HLA命名委员会正式命名.
목적 연구인류백세포항원(human leukocyte antigen,HLA)신등위기인HLA_B*9536화B*4612적분자궤제.방법 채용Invitrogen추제시제합추제표본DNA,이용단련특이성인물PCR방법확증양본HLA-B기인제2～4외현자,대PCR산물직접진행HLA-B기인제2、3、4외현자쌍향측서분석.결과 선증자표본존재2개HLA-B등위기인,경HLA Blast험증균위신적등위기인,신적등위기인서렬이체교GenBank(EU081878화EU081879),경세계위생조직HLA명명위원회정식명명위HLA-B*9536화HLA-B*4612.HLA-B*9536제2～4외현자서렬여최접근적B*1505상비,재제3외현자존재일개감기적불동,즉제544위G→A개변,도치제158위안기산Ala→Thr;HLA-B*4612제2～4외현자서렬여최접근적B*4601상비,재제3외현자존재일개감기적불동,즉제363위G→C,도치제97위안기산Arg→Ser.결론 재동일표본중발현량개신적HLA-B등위기인,병피세계위생조직HLA명명위원회정식명명.
Objective To identify two novel HLA alleles HLA-B*9536 and B*4612,in an individual.Methods DNA was extracted from whole blood by Invitrogen DNA extraction kit.The amplification for HLA-B exons 2-4 of the proband was performed separately with allele group specific primers and the PCR products were directly sequenced for exons 2-4 in both direction.Results There were two novel HLA-B alleles in the proband.The sequences of the two alleles have been submitted to GenBank (EU081878 and EU081879).The two alleles have been officially named as B*9536 and B*4612 by the WHO Nomenclature Committee.The sequence of exons 2-4 of HLA-B*9536 showed one nucleotide difference in exon 3 at position 544(G to A)comparing with the closest allele B*1505,which resulted in an amino acid change from Ala to Thr at codon 158.In the HLA-B*4612 allele,there was one nucleotide change in exon 3 at position 363(G to A),when compared to the closest allele B*4601,which lead to an amino acid change from Arg to Ser at codon 97.Conclusion Two novel HLA-B alleles were identified in one individual and have been officially named by the WHO Nomenclature Committee.