Cloning and Bioinformatics Analysis of TLR2 cDNA in Buffalo

In this report,Toll-like receptor 2(TLR2) cDNA of buffalo was amplified partly by RT-PCR.The amplified fragments were spliced into a full-length fragment through the overlapping areas.The splicing product was then cloned into the pMD20-T vector,the recombinant plasmid was confirmed by PCR and endonuclease digestion,then it was sequenced and bioinformatically analyzed.Results showed that TLR2 cDNA was 2 455 bp in length and contained a 2 355 bp ORF,encoding 784 amino acids,with the molecular weight of 90.2 ku,and the theoretical IP was 6.92.The nucleotide sequence of the cloned TLR2 ORF shared 99.53% homologies with that of buffalo published in GenBank.And the homology between the nucleotide sequence of the cloned TLR2 ORF and that of cattle,horse,human,chimpanzee and dog were 97.88%,84.94%,83.12%,82.95% and 82.74% respectively.These data suggested that TLR2 gene was relatively conserved among different species.TLR2 is a transmembrane protein,contains several domains like LRR,LRR-TYP,LRRCT and TIR,which are consistent with the common structure of Toll-like receptor family members.The successful clone,sequence analysis and structure prediction of TLR2 cDNA of buffalo lays a foundation for further research on the biological function of the gene.