DGGE技术检测小核核糖核蛋白多肽N基因rs220030位点的多态性

Polymorphism Study of Small Nuclear Ribonucleoprotein Polypeptide N Gene rs220030 by DGGE

目的分析小核核糖核蛋白多肽N(small nuclear ribonucleoprotein polypeptide N,SNRPN)基因SNP位点rs220030在中国汉族人群中的基因结构特征及多态性,获得群体遗传学资料。方法应用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)技术对100例上海地区汉族健康无关个体SNRPN基因上游启动子区域内的SNP位点rs220030进行检测,该位点TaqMan SNP的基因分型被用来确定在某些典型样品的DGGE的结果。采用TaqMan探针技术进行验证。结果 rs220030位点经DGGE检测,杂合子CT检出4条谱带,CC、TT纯合子检出1条谱带,TaqMan探针分型结果与DGGE分型结果一致。100例上海地区汉族人群rs220030位点共检出CC 34例,CT 41例,TT 25例;等位基因C和T的频率分别为0.545和0.455,H为0.500,PIC为0.373,DP为0.654,PE为0.186,样本群体基因型频率符合Hardy-Weinberg平衡。结论 DGGE技术适用于小样本群体的低通量SNP位点多态性分析,SNRPN基因rs220030位点参数的多态性信息含量能够满足目前法医遗传学亲权鉴定和个人识别的需要。

Objective To analyze the polymorphism of rs220030,a SNP which is located in the promoter region of small nuclear ribonucleoprotein polypeptide N（SNRPN） gene in the Chinese Han population and to obtain the data of population genetics.Methods The denaturing gradient gel electrophoresis（DGGE） method was applied to detect the polymorphism of rs220030 in 100 unrelated and healthy individuals from the Shanghai Han population.The genotyping result of this SNP was confirmed by TaqMan assay in some typical samples.Results DGGE results showed 4 bands for CT heterozygote,and 1 band for CC or TT homozygote,and those results were confirmed by The TaqMan SNP genotyping assays.Genotyping results showed 34 individuals with CC,41 with CT and 25 with TT of rs220030.The allele frequencies for C and T were 0.545 and 0.455,respectively.H was 0.500,PIC was 0.373,DP was 0.654,and PE was 0.186.The distribution of genotype frequencies were in Hardy-Weinberg equilibrium.Conclusion DGGE is a quick and effective method in the analysis of SNP polymorphism in small population.Statistical parameters of rs220030 for forensic evaluation meet the requirements for forensic identification and paternity testing.