Cloning,Expression and Identification of CD14 cDNA of Buffalo

Objective:This study was aimed to clone and express bCD14(buffalo cluster of differentiation antigen 14),then carry out Western Blot for it.Method:RT-PCR method were used to clone CDS area of cluster of bCD14,then,the PCR products of bCD14 gene was cloned into the expression vector PET-28a,and transformed into the host strain E.coli BL21.After the inducible expression by IPTG,the target protein was analyzed under denaturing condition,Western blot were used to identify the fusion protein.Result:The results showed that bCD14 gene contain a 1122 bp ORF encoding 373 amino acids,compared with Indian buffalo,Norway rat and human,the homology of cDNA were 97.95%,68.78% and 78.60%;the homology of amino acid were 96.78%,61.27% and 72.34%,then the 46kD target protein was confirmed by Western blot.Conclusion:The CDS area of bCD14 was successfully cloned,expressed,which laid a solid foundation for the in-depth function study of bCD14.