非探针法高分辨率熔解曲线(HRM)对FecB基因分型方法的研究

Study on a genotyping method of the FecB gene by non-probe high-resolution melting(HRM) curve

为了建立一种基于非探针法高分辨率熔解曲线(HRM)技术对绵羊FecB基因高效分型的方法,试验选用健康、成年绵羊48只,利用HRM和限制性片段长度多态性(PCR-RLFP)对绵羊多胎主效基因FecB基因进行分型效果研究。结果表明:FecB基因具有3种基因型,分别是野生型(AA)37.50%、杂合型(GA)33.33%、纯合突变型(GG)29.17%;通过对基因测序比对、HRM和PCRRLFP结果的准确率比较,HRM的准确率达到了95.83%(46/48),高于RLFP的分型结果,所需试验时间大大缩短。说明非探针法HRM技术是一种高效率、节省人工的FecB基因分型方法。

To establish a highly efficient method for genotyping the FecB gene of sheep based on non -probe high -resolution melting （HRM） technology, 48 healthy adult sheep were used in the test. HRM and polymerase chain reaction - restriction fragment length polymorphism （ PCR - RFLP） were used to study the genatyping results of the FecB gene as a main effect gene of multifetal sheep. The results showed that there were three types of genotypes in the FecB gene, including 37.50% of wild type （ AA）, 33.33% of heterozygous type （ GA）, and 29.17% of homozygous mutant type （GG）. The accuracy of the genotyping result by HRM reached 95.83% （46/48）, which was higher than that by RLFP and the testing time was greatly shortened through gene sequencing and comparing the accuracy of the genotyping results between HRM and RLFP. The results indicate that non - probe HRM technique is a high - efficiency and saving - labor genotyping method of the FeeB gene.