A research on different ways of recombinant to express caf1M protein of Yersinia pestis

Objective Different methods to clone and express the existential state and immunological activities of the caf1M protein of Yersinia pestis.Methods There different vectors were selected to express caf1M,pET32a(+),pGEX4t-1 and pGBTNH.caf1M's immunological activities were evaluated by anti-Yersinia pestis EV rabbit sera.Results Four recombinant plasmids were successfully constructed.Three plasmids were constructed by removing the coding sequence of the signal peptide from caf1M and connecting it with one of the vector plasmids pET32a(+),pGEX4t-1 or pGBTNH.The fourth plasmid was constructed by connecting the complete caf1M with the vector plasmid pGEX4t-1.The four plasmids transformed into E.coli display high expression of the 4 recombinant proteins.Soluble recombinant protein was expressed by recombinant plasmid of the gene of caf1M removing the coding sequence of signal peptide connecting with vector plasmid pGEX4t-1,the other three recombinant proteins are in form of inclusion bodies.Besides the recombinant plasmid containing pGBTNH,the three other recombinant proteins reacted with anti-Yersinia pestis EV76 rabbit sera.Conclusion Four types of recombinant caf1M protein were successful expressed.Three out of the four recombinant proteins displayed idiosyncratic reaction.Thus,signal peptides and expressing vectors had an impact on the expression of recombinant proteins.