猪CAPN1基因部分外显子及3′UTR区的SNPs检测

以野猪、民猪和大白猪为研究对象, 根据网上公布的序列设计了7对引物, 采用测序、PCR-SSCP和PCR-RFLP方法对CAPN1基因的部分外显子和3′UTR区进行了单核苷酸多态性检测和基因型分析, 探讨CAPN1基因多态性与瘦肉率和嫩度的关系。研究发现11个SNPs, 其中5个位于外显子, 4个位于内含子, 2个位于3′UTR区, 外显子中的突变有一处是错义突变, 导致了蛋白质多肽链第260位氨基酸发生了M/V的替代。群体遗传学分析表明, 在所检测的各多态位点上, 野猪、民猪、大白猪3个品种间不同基因型的分布都存在着极显著的差异(P<0.01), 而野猪和民猪之间各基因型的分布差异不显著(P>0.05), 民猪和大白猪之间各基因型的分布存在着极显著的差异(P<0.01)。结合品种特性分析表明, P4、P6引物和3′ UTR区HinfⅠ位点所检测的不同基因型和瘦肉率具有一定的相关性。

Analysis of SNPs in partial exons and 3' UTR of CAPN1 in porcine

In this study, Wild boar, Min pig and Yorkshire were used to investigate the effect of CAPN1 gene on lean meat percentage and tenderness. Seven pairs of primers for exons and 3'untranslated region of CAPN1 gene were designed according to pig sequences from GenBank. Polymorphisms within exons were detected by PCR-SSCP method. SNPs in 3'untranslated region were detected by DNA sequencing, and PCR-RFLP method was then developed to screen the population. Totally, 11 SNPs, of which 5 from exons including one missense mutation resulting in amino acid substitution of M/V at the position of 260, 4 from introns and 2 from 3'untranslated region, were identified. Population genetics analysis showed that the distribution of genotypes among Yorkshire, Min pig and Wild boar were extremely significant difference (P<0.01), while there were no significant difference between Min pig and Wild boar (P>0.05), and extremely significant difference existed between Min pig and Yorkshire (P<0.01). Association of polymorphisms with breed characteristics indicated that the mutations detected by P4, P6 primers and that resulting in the change of Hinf 1 recognition site may be related to lean meat percentage.