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作 者:蒋伟宏[1] 邹嫣琼[1] 林标扬[1] 陆嫣[1] 费虹明[1] 陈仁彪[1]
机构地区:[1]上海第二医科大学生物学教研室
出 处:《中华微生物学和免疫学杂志》1999年第2期156-157,共2页Chinese Journal of Microbiology and Immunology
基 金:国家自然科学基金
摘 要:目的对在用PCR/SSO方法作我国彝族群体样本HLAD区寡核苷酸分型中发现的1例DRB1的杂合子进行等位基因序列分析。方法用DRB基因类引物扩增该特殊的DRB1杂合子细胞基因组的DRB基因的第二外显子。扩增产物经纯化转染大肠杆菌JM101,克隆后再经酚/氯仿抽提后得到单链M13DNA。利用ABI377测序仪自动测序。结果彝族这一特殊的DRB1杂合子细胞的DRB1基因阳性克隆,经序列分析证实了该DRB1杂合子中一个等位基因确为DRB1*1202,与PCR/SSO分型一致;另一个新等位基因DRB1*15Y2(暂定)等位基因在第二外显子的47位由编码苯丙氨酸的TTC(DRB1*1502)变为编码酪氨酸的TAC。结论在云南彝族样本中发现的DRB1*15Y2,其47位由TAC(酪氨酸)置换了相应于DRB1*1502的TTC(苯丙氨酸)。在我国这样一个多民族国家中,就HLA系统而言,可能还有更多新等位基因有待鉴定。Objective One special DRB1 heterozygote which was found in oligotyping of HLA D region in Chinese Yi population by using PCR/SSO method, was to be sequenced for the gene of the allele. Methods Amplification of exon 2 of DRB gene in this heterozygote was carried out with genomic DNA templet and generic primers and the amplicon was cloned in M13 for further sequencing. Results One allele was identified as DRB1*1202, another allele whose PCR/SSO hybridization pattern was similar to but different from DRB1*1502 was considered as DRB1*15 variant. Analysis of nucleotide sequence revealed that this allelic variant differed from DRB1*1502 at codon 47 (TAC instead of TTC), resulting in the substitution of a tyrosine for phenylalanine in the DRβ chain. Conclusion The DRB1 heterozygote allele gene could be nominated as DRB1*15Y2 temporarily.
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