消化后片段长度差异等位基因特异性PCR技术——印记SNP rs220028多态性和甲基化模式研究  

The multiplex analysis of epigenetic markers and genetic markers by post-digestion mutagenically separated PCR

作　　者：赵贵森[1] 黄代新[1] 冯文芳[2] 杨庆恩[1] 

机构地区：[1]华中科技大学同济医学院法医学系,武汉430030 [2]华中科技大学同济医学院生物化学与分子生物学系,武汉430030

出　　处：《中华医学遗传学杂志》2005年第1期58-60,共3页Chinese Journal of Medical Genetics

摘　　要：目的建立一种能同时分析甲基化和单核苷酸多态性(singlenucleotidepolymorphism,SNP)的新方法。方法以印记SNPrs220028(A/G)为例,基因组DNA经甲基化敏感限制酶消化后用片段长度差异等位基因特异性PCR(fragmentlengthdiscrepantallele-specificPCR,FLDAS-PCR)分型;用扩增产物酶消化法来排除酶切位点序列变异,证实检出的是甲基化等位基因。结果消化后FLDAS-PCR可选择性检出甲基化等位基因,家系分析表明其来自母亲。等位基因频率A=0.5085,G=0.4915,多态信息含量为0.3749。结论消化后FLDAS-PCR技术可以实现甲基化和SNP的复合分析;印记SNPrs220028在Prader-Willi/Angelman综合征诊断中有潜在意义。Objective To establish a novel method for the multiplex analysis of the methylation and single nucleotide polymorphism (SNP). Methods The imprinted SNP rs220028 was chosen as a model. Genomic DNA, after being digested with methylation sensitive restriction enzyme, were typed by mutagenically separated PCR (MS-PCR). The polymorphism of restriction site was excluded by PCR-RFLP. Results By post-digestion MS-PCR, the methylated allele was detected selectively, the maternal origin of which was confirmed by pedigree analysis; A=0.5085,G=0.4915,PIC=0.3749. Conclusion The multiplex analysis of methylation markers and SNP can be achieved by post-digestion MS-PCR. The imprinted SNP locus rs220028 is a potentially useful marker in screening Prader-Willi/Angelman syndrome.

关 键 词：甲基化 SNP 等位基因 检出 特异性 PCR技术 诊断 酶切位点 片段 PCR) 

分 类 号：Q78[生物学—分子生物学]