中国草原红牛CAPNI基因的遗传多样性分析

Analysis on the Genetic Diversity of CAPN IGene in China Red Steppe Cattle

[目的]利用PCR-SSCP技术研究中国草原红牛CAPN I基因的多态性。[方法]以90头草原红牛的全血为试材,用酚/氯仿/异戊醇法提取基因组DNA,采用PCR-SSCP技术分析了钙蛋白酶I基因(CAPN I)在草原红牛群体中的多态性。[结果]试验所设计的18对引物用于PCR扩增均获得良好的结果。经PCR-SSCP分析,E7-1引物扩增的群体出现了AA、BB、AB和AC 4种基因型;E14-1引物扩增的群体出现了AA和BB 2种基因型,E14-5、E14-6和E20-2引物扩增的群体出现了AA、BB和AB 3种基因型。对不同基因型的测序表明,在第14内含子4 685 bp处发生了C-T的碱基突变,在第17内含子6 545 bp处发生了C-T的碱基突变,在第21内含子上8 462 bp处发生了G-A、8 561 bp处A-G、8 696 bp处A-T的碱基突变。[结论]该研究为改善牛肉质性状的分子育种提供新的分子生物学基础数据。

[ Objective ] The aim was to research the polymorphism of polymorphism of calpain I（ CAPN I） gene in China Red Steppe by using PCR-SSCP technique. [ Method] With the whole blood of 90 head China Red Steppe as the tested material, the genome DNA was extracted by using phenol/chlorofonn/isoamyl alcohol method and the CAPN I gene in the China Red Steppe group by using PCR-SSCP technique. [ Resuit] The designed 18 pairs of primers used to PCR amplification all got the better results. Through PCR-SSCP analysis, the group of E7-1 primer amplification appeared 4 kinds of genotypes of AA, BB, AB and AC ; the group of El4-1 primer amplification appeared 2 kinds of genotypes of AA and BB, the group of E14-5, E14-6, E20-2 primer amplification appeared 2 kinds of genotypes of AA, BB and AB. The sequencing on different genotypes showed that the base mutation of C-T was happened at the 14th intron 4 685 bp, the base mutation of C-T was happened at the 17th intron 6 545 bp, the base mutation of G-A, A-G, A-T were happened at the 21th intron 8 462, 8 561 and 8 696 bp resp. [ Conclusion] This research provided the basic data of molecular biology for the molecular breeding of improving the beef quality characteristics.