非标记探针HRM法在中国对虾EST-SNP筛选中的应用

Application of unlabeled probe by HRM in development of EST-SNPs in Fenneropenaeus chinesis

利用高分辨率溶解曲线(High resolution melt,HRM),结合非标记探针技术,对中国对虾转录组EST测序序列中的118个候选SNP位点进行了位点多态性检测,获得了39个具有二等位基因多态性的SNP位点,占总候选位点的比例为33.1%。对这些SNP位点在一个48尾中国对虾群体中的多态性进行了分析,结果表明,观测杂合度Ho和期望杂合度He的分布范围分别为0.000～0.947和0.049～0.506,有效等位基因数分布范围为1.051～1.999,平均为1.574;多态信息含量分析显示39个位点的PIC值范围为0.0476～0.375,平均为0.272。另外,基因功能注释表明,39个多态SNP所在contig序列所对应的基因大都与免疫相关。以上这些结果表明,非标记探针HRM是一种简单快速而有效的SNP开发技术,可为中国对虾群体遗传学和遗传育种分析提供有效的候选标记。

In Fenneropenaeus chinesis,118SNPs were selected for validation using high resolution melting（HRM）assays with unlabeled probes,in which 39（33.1%）loci were polymorphic.Polymorphism of these 39distinct loci was assessed using 48individuals,and the results showed that the values of N e,H o,H e and PICvaried from 1.051 2to 1.999,from 0.000 to 0.947,from 0.049to 0.506,from 0.049to 0.506,respectively.The 39contigs,to which 39 loci belonged,were annotated with Nr database.The annotation showed that,most corresponding genes were associated with immunity.The results indicated that unlabeled HRM is a simple,rapid and efficient method for SNP genotyping.These polymorphic markers will be useful genetic tools in the population genetics and breeding programs for Fenneropenaeus chinensis.