四引物扩增受阻突变体系PCR技术在中国明对虾SNP基因分型中的研究  

Tetra-primer amplification refractory mutation system-polymerase chain reaction in SNP genotyping of shrimp Fenneropenaeus chinensis

作　　者：张建勇[1,2,3] 王清印[2] 王伟继[2] 孟宪红[2] 孔杰[2] 张全启[1] 

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266002 [2]中国水产科学研究院黄海水产研究所,山东青岛266071 [3]山东理工大学生命科学学院,山东淄博255049

出　　处：《中国水产科学》2011年第4期751-759,共9页Journal of Fishery Sciences of China

基　　金：国家自然科学基金资助项目(31072206);山东省自然科学基金项目资助(ZR2009DQ002)

摘　　要：采用四引物扩增受阻突变体系PCR（Tetra-primer ARMS-PCR）技术,对中国明对虾（Fenneropenaeus chinensis）的80个单核苷酸多态性位点（Single Nucleotide Polymorphisms,SNPs）进行验证。结果表明,通过优化PCR反应条件、调整内外引物浓度和采取Touchdown PCR程序可优化扩增效果,80组引物中有20组引物得到良好的分型效果。群体多态性分析表明有效等位基因数（Ne）、观测杂合度（Ho）、期望杂合度（He）及多态性信息含量（PIC）的范围分别为1.127~1.993、0.136~0.607、0.119~0.492和0.145~0.373。结果显示四引物扩增受阻突变体系技术聚合酶链式反应是一种简单快速而有效SNP基因分型的方法。Tetra-primer amplification refractory mutation system-polymerase chain reaction（Tetra-primer ARMA-PCR） was introduced to investigate single nucleotide polymorphisms（SNPs） genotyping in Chinese shrimp（Fenneropenaeus chinensis） and 80 putative SNPs loci were studied.Twenty out of the 80 SNP tetra-primer ARMA-PCR primer sets were validated following touchdown profiles and the outer and the expected inner bands were amplified.Homozygous and heterozygous were detected by agarose gel and the genotypes were obtained.Polymorphism of these distinct loci was assessed using 30 individuals,and the results showed that the 20 loci were all polymorphic.The values of Ne,Ho,He and PIC varied from 1.127 to 1.993,from 0.136 to 0.607,from 0.119 to 0.492 and from 0.145 to 0.3730,respectively.The results indicated that tetra-primer ARMA-PCR is a simple,rapid and efficient method for SNP genotyping which make it useful in a broad aspects of Fenneropenaeus chinensis genetic and breeding studies.

关 键 词：单核苷酸多态性 基因分型 中国明对虾 四引物扩增受阻突变体系PCR 

分 类 号：S968[农业科学—水产养殖]