H19基因上游差异性甲基化区SNPs多态性研究

目的获得H19基因上游差异性甲基化区中SNPs的群体遗传学信息。方法采用PCR和测序技术,对105例中国北方汉族健康无关个体H19上游启动子区检测;使用Haploview 4.1和PowerStats V12软件进行统计学分析。选用甲基化敏感的限制内切酶(msRE)HpaⅡ,检测5个家系样本H19等位基因的亲代来源。结果测序结果显示,H19启动子区含有13个SNPs,组成5种单倍型,13种单倍型组合,其个体识别能力为0.856、多态性信息含量为0.67、非父排除率为0.498。经msRE HpaⅡ消化母源等位基因后,进行PCR及测序分析,检测出父源等位基因,排除1例和肯定4例家系的亲缘关系。结论 DNA甲基化标记和SNPs多态性检测,可同时进行多态性分型并确定等位基因的亲代来源,具有较高的法医学应用价值。

A study on SNP polymorphisms in the differentially methylated region upstream of the human H19 gene

Objective To obtain the population genetics data of SNPs in the differentially methylated region upstream of the human H19 gene.Methods By means of PCR and sequencing,the promoter region upstream of the H19 gene were investigated from total 105 unrelated Chinese Han individuals;Haploview 4.1 and PowerStats V12 were used for the statistical analysis.The parental alleles of the 5 families were detected after digested with the methylation-sensitive restriction enzyme(msRE) HpaⅡ.Results Sequencing results showed that there were 13 SNPs in the promoter region of the H19 gene.Five haplotypes and thirteen phenotypes were observed in 105 unrelated individuals,and the power of discrimination(DP),polymorphism information content(PIC) and probability of paternity exclusion(PE) were 0.856,0.67 and 0.498 respectively.The paternal alleles were detected by the amplification and the sequencing after digested with msRE HpaⅡ,the paternity was excluded in one case and others were confirmed.Conclusion Combined with the SNPs,the parentally imprinted allele(PIA) typing method can discriminate the parental origin of alleles and be used in forensic science as a valuable tool.