野猪、家猪及野家杂种猪CAPN1基因3′UTR区的SNPs检测及PCR-RFLP分析

The SNPs detection and PCR-RFLP analysis on 3′UTR of CAPN1 gene in wild boar,swine and hybrid swine

μ-钙蛋白酶(μ-calpain,CAPN1)是动物体内肌原纤维蛋白降解的主要酶,为了揭示其基因变异与瘦肉率和嫩度的关系,文章以野猪、家猪及野家杂种猪为研究对象,通过测序和PCR-RFLP方法对CAPN1基因3′UTR进行了SNPs检测和多态性分析。结果表明,共发现7个SNPs,分别是C114T,G220C,C344T,T380C,G501A,T543C,T610C;对G220C/T380C两处突变建立了基于限制性内切酶HinfⅠ/AlwNⅠ的PCR-RFLP检测技术;群体遗传学分析表明,在这两个位点不同基因型在不同品种(品系)猪中的分布都存在着极显著的差异(P<0.01),野猪和民猪之间的差异都不显著(P>0.05),并且HinfⅠ位点的B等位基因频率具有随着品种瘦肉率的增加而增多的趋势。该结果为寻找高瘦肉率的分子标记提供了基础。

μ-calpain(CAPN1) is the major proteolysis of muscle fibers.In order to further reveal the relationship between the variation of CAPN1 gene and the lean meat percentage and tenderness,the SNPs in 3′UTR of CAPN1 were detected by sequencing method.Seven SNPs,which were C114T,G220C,C344T,T380C,G501A,T543C,T610C respectively,were found.PCR-RFLP method was then developed to screen the wild boar,Min pig,Yorkshire,Duroc and hybrid swine in G220C and T380C sites based on the restriction endonuclease enzyme HinfⅠ/AlwNⅠ.The population genetics analyses showed that distribution of genotypes among these five breeds(lines) were extremely significant differences(P<0.01) in the two mutation sites,while there were no significant differences between Min pig and wild boar(P>0.05).The gene frequency of allele B in HinfⅠmutation site increased with average lean meat percentage of breed.The results lay foundation for selection of molecular marker of high meat percentage.