Zhu Fa-ming, Zhang Wei, Lu Qin-feng, He Ji, Wang Wei, Han Zhe-dong, Yan Li-xing
Blood Center of Zhejiang Province, Hangzhou, Zhejiang 310006, P.R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2006 Feb;23(1):47-9.
To investigate the molecular genetics basis for HLA novel allele HLA-DRB1*1212 in Chinese population.
Genomic DNA was extracted from whole blood by salting-out method. HLA-DRB1 gene exon 2 was amplified by PCR with group-specific primers from genomic DNA. PCR products were cut back from agarose gels and purified to sequence directly. The polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSO) was performed to confirm the mutations which were detected by sequencing in this study.
The sequencing results showed HLA-DRB1 alleles of the proband as DRB1090102 and the novel allele. The sequences of the novel allele have been submitted to GenBank (AY899825). Through BLAST analysis, the novel allele was found to be different from DRB1120101 at position 199A-->C in exon 2, that results in an amino acid change from Ile to Leu at codon 67.
This allele is a novel and has been officially named as DRB1*1212 by the WHO Nomenclature Committee.
研究中国人群中HLA新等位基因HLA - DRB1*1212的分子遗传学基础。
采用盐析法从全血中提取基因组DNA。用组特异性引物通过PCR从基因组DNA中扩增HLA - DRB1基因外显子2。PCR产物从琼脂糖凝胶中切下并纯化后直接测序。进行聚合酶链反应 - 序列特异性寡核苷酸探针（PCR - SSO）检测以确认本研究中测序检测到的突变。
测序结果显示先证者的HLA - DRB1等位基因为DRB1090102和新等位基因。新等位基因的序列已提交至GenBank（AY899825）。通过BLAST分析，发现该新等位基因在外显子2的第199位A→C处与DRB1120101不同，导致第67位密码子的氨基酸从异亮氨酸变为亮氨酸。
该等位基因是一个新基因，已被世界卫生组织命名委员会正式命名为DRB1*1212。
