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目的 研究中国汉族人群Ax亚型的分子生物学特点。方法 首先采用双重PCR RLFP方法 ,对 8份血清学试验疑为Ax及AxB的标本 ,做ABO初步基因分型 ,明确其中的A、B或O基因 ;然后用限制性内切酶mobI筛选ABO基因第 7外显子中的T6 4 6A的突变。对不具有该突变而血清学疑为Ax的标本以及血清学和基因分型不一致的标本 ,则进一步对其ABO基因第 6和 7外显子做深入的克隆测序分析。结果 8份标本中有半数含有T6 4 6A点突变 ,剩余 4份中 ,1份为包含C4 0 7T和C4 6 7T错义突变的A weak 0 1等位基因 ,1份为携带新的核苷酸变异的Ax (C4 6 7T和C74 5T)等位基因 ,另外 2份血清学疑为AxB ,初步基因分型为BO的个体 ,克隆测序的结果显示两者均有正常的O基因 ,但是其B等位基因均发生了新的nt6 4 0位A G突变。结论 在中国汉族人群中检测到新的Ax及B(A)等位基因
Objective To study the molecular characteristics of Ax subtypes in Chinese Han population. Methods The double-stranded PCR (RLFP) method was used to detect the ABO genotypes in eight serological samples suspected to be Ax and AxB, and to identify the A, B or O genes. The ABO gene was then screened with restriction endonuclease mobI Mutation of T6 4 6A in exon 7. For those specimens that did not have this mutation and were suspected to be suspected of Ax, and serological and genotyping inconsistencies, further in silico cloning and sequencing analyzes of exons 6 and 7 of the ABO gene were performed. Results One half of 8 specimens contained T6 4 6A point mutations and the other 4 were A weak 0 1 alleles containing C4 0 7T and C4 6 7T missense mutations. The acid-variant Ax (C4 6 7T and C74 5T) alleles, and the other 2 individuals suspected of being AxB serologically, with a preliminary genotype of BO, were cloned and sequenced and both showed normal O genes, A new nt640 AG mutation occurred in the B allele. Conclusions New Ax and B (A) alleles were detected in Chinese Han population