荧光-构象敏感凝胶电泳技术筛查上海市郊区野生小家鼠线粒体编码区SNP位点

利用通用引物荧光PCR方案, 应用荧光-构象敏感凝胶电泳(fluorescence-based conformation sensitive gel elec-trophoresis, F-CSGE)和DNA直接测序分型技术, 对上海地区64只野生小家鼠线粒体DNA(mtDNA)编码区进行序列分析, 在上海市郊区野生小家鼠群体中初步检测mtDNA编码区SNP, 以发现合适的遗传位标用于野生小家鼠遗传多态性分析。结果发现: F-CSGE所有存在SNP突变的峰图中同源双链和异源双链峰电泳泳动距离差异均较为明显, 在检测未知SNP中无假阳性出现, 检测效率高。F-CSGE检出野生小鼠mtDNA编码区SNP 24个, 其中新发现SNP为16个。结果表明, F-CSGE可用于mtDNA编码区SNP检测, 新发现的SNP可作为遗传位标用以研究整个上海地区野生小家鼠的遗传结构和遗传多态性。

SNP discovery by F-CSGE in the coding region of mitochondrial DNA in wild house mice from Shanghai suburb

This study was performed to discover SNPs for genetic polymorphism analysis of mitochondrial DNA from wild house mice. Universal primer florescent PCR, fluorescence-based conformation sensitive gel electrophoresis (F-CSGE) and DNA sequencing were conducted to analyze the coding region of mitochondrial DNA. Different types of unknown mutations were recorded by variable F-CSGE patterns without false positive. Twenty-four SNPs, sixteen of which were first discovered in the coding region of mitochondrial DNA, were found in 64 wild house mice from 4 districts in Shanghai. Therefore, F-CSGE was proved to be powerful technique for SNP discovery in the coding region mitochondrial DNA. The novel SNPs can be used as molecular markers to analyze population structure and genetic polymorphisms of the wild house mice in Shanghai.