猪STCH基因的Bi-PASA遗传标记及多态性研究

微粒体应激 70蛋白三磷酸腺苷酶 (STCH)基因属于应激 70蛋白基因伴侣家族 ,在机体免疫反应和疾病抵抗力等方面起重要作用。根据人和小鼠STCH基因的保守序列设计引物 ,PCR扩增到猪STCH基因第5外显子 4 4 5bp片段。序列测定显示 ,猪STCH基因与人和小鼠STCH基因分别具有 87 13%和 80 4 5 %的同源性。通过测定和比较中国梅山猪、欧洲约克夏猪及PIC商品猪的STCH基因序列 ,发现在猪STCH基因编码区第 5外显子 10 5 0位点上存在一个单碱基突变位点。利用双向特定等位基因PCR扩增法 (Bi PASA)建立了检测猪STCH基因变异的遗传标记 ,并用该标记分析了STCH基因在中国家猪 (梅山猪、荣昌猪和金华猪 )、欧洲家猪 (约克夏猪、大白猪 )、商品猪 (PIC合成系 )以及欧洲野猪的基因频率和多态性。本研究建立的Bi PASA遗传标记和基因变异信息 ,将为进一步分析猪STCH基因变异与经济性状的相关分析提供基础资料。

Bi-PASA Genetic Marker and Polymorphism of the Porcine STCH Gene

Microsomal stress 70 protein ATPase core (STCH) is a member of the stress 70 protein chaperone family and plays a major role in the immune response and disease resistance of host.Based on the conserved regions of STCH gene sequences from human and mouse,a pair of primers was designed for PCR amplification of porcine STCH gene.A fragment of 445 base pairs product in the exon 5 of pig STCH gene was successfully amplified with the primers.The porcine STCH sequence determined from this fragment showed 87.13% and 80.45% nucleotide identity to the corresponding human and mouse STCH sequences,respectively.A single nucleotide polymorphism (SNP) was found at the position 1 050 of coding sequence in the exon 5 of porcine STCH gene according to the sequencing and alignment of STCH genes from Meishan pigs,Yorkshire pigs and PIC commercial pigs.As no cutting sites of all known restriction enzymes could be found for the SNP,Bi-directional PCR amplification of specific alleles (Bi-PASA) was employed to detect the mutation.According to the porcine STCH Bi-PASA marker,the gene allele frequencies and gene polymorphisms were analyzed in the Chinese pig populations (Meishan pigs,Rongchang pigs,Jinhua pigs) and European populations (Yorkshire pigs,Large white pigs) as well as a commercial line (PIC pigs).The STCH Bi-PASA marker established in this study and polymorphic information obtained from population analysis may be helpful for the further association study between the pig STCH gene mutations and economic traits in pigs.