中华鳖(Pelodiscus sinensis)不同品系线粒体SNP的分型与鉴定

IDENTIFICATION OF DIFFERENT STRAINS OF PELODISCUS SINENSIS BY USING HIGH RESOLUTION MELTING(HRM) ANALYSIS OF SNPS IN MITOCHONDRIAL DNA

通过对四个中华鳖(Pelodiscus sinensis)养殖品系(太湖品系、台湾品系、日本品系、黄河品系)的线粒体部分序列进行测序对比,筛选出不同品系中华鳖的品系特异性SNP位点,并设计特异性引物,利用高分辨率熔解曲线法进行SNP分型与鉴定。结果表明,所设计的7对引物组均可扩增出80—150bp大小不等的中华鳖线粒体序列,并成功用于中华鳖的SNP分型。在这7个位点中,其中可鉴定中华鳖日本品系和太湖品系的各1个,可鉴定台湾品系的有2个,可鉴定黄河品系的有3个。此方法能够快速、简便地鉴别以上四个中华鳖品系,为鉴别市场中出现的假冒鳖,并指导品种选育工作提供技术支持。

Partial sequences of mitochondrial DNA from four Pelodiscus sinensis strains(Taihu Lake, Taiwan, Japanese, and Huanghe River strains) were amplified and sequenced to locate strain-specific SNPs. Corresponding primers were designed and high-resolution melting(HRM) technique employed for genotyping these SNPs. Seven SNPs were detected by HRM, whose length of amplified DNA sequence ranged from 80bp to 150bp, of which one was the Taihu Lake strain specific, two specific to the Taiwan strain, one to Japanese strain, and three to the Huanghe River strain. This method is fast and simple, and may offer a technical support to the strain identification and breeding of P. sinensis in practice.