猪TLR2基因Bi-PASA遗传标记的建立及多态性研究

根据人和小鼠TLR2基因序列设计了特异性PCR引物,优化PCR条件后扩增到中国梅山猪、欧洲约克夏猪、PIC-2系及PIC-3系商品猪的TLR2基因690 bp的基因片段,并对其进行序列分析。序列分析结果显示,猪TLR2基因多态性程度低,发现在猪TLR2基因编码区第1 255位点上存在1个单碱基突变位点。利用双向特定等位基因PCR扩增法(B i-PASA)建立了检测猪TLR2基因变异的遗传标记。利用猪TLR2基因的B i-PASA标记,分析了TLR2基因在大河乌猪、撒坝猪和黔邵花猪中的基因频率和多态性。研究建立的B i-PASA遗传标记和基因变异信息,将为进一步分析猪TLR2基因变异与疾病抵抗力及经济性状的相关分析提供基础资料。

Bi-PASA genetic marker and polymorphism of the porcine TLR2 gene

TLR2 gene is an important member in the TLR(Tolllike receptors)genes family,which play an important role in the recognition of pathogen especially in innate immune system.TLR gene mutations are important genetic bases on the difference of disease-resistance in defence response to pathogens.In this study,we designed specific pig primers in term of human's and mouse's TLR2 gene's reported sequences in the Genbank.After optimizing the conditions of polymerase chain reaction(PCR),we successfully amplified 690 bp of TLR2 gene of Meishan,Yorkshire,PIC-line2 and PIC-line3,and purified and sequenced the PCR products.Sequence analysis showed that porcine TLR2 gene had little polymorphisms.In the Sequence,only one single nucleotide polymorphism(SNP) was found at the position 1 255 of coding sequence of the PCR products according to the sequencing and alignment of TLR2 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-PASA(Bi-directional PCR amplification of specific alleles) was employed to detect the mutation.According to the porcine TLR2 Bi-PASA genetic marker,the gene allele frequencies and gene polymorphisms were analyzed in the populations of Dahe black pigs,Saba pigs and Qianshao spotted pigs.The TLR2 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 TLR2 gene mutations and economic traits in pigs.