Construction of recombinant prokaryotic plasmid and expression of cathepsin B endopeptidase of Schistosoma japonicum

Objective To discuss the construction of recombinant prokaryotic expression plasmid Sjcb2/pWR450-1 and expression cathepsin B endopeptidase of Schistosoma japonicum in E.coli DH5α. Methods The gene of cathepsin B endopeptidase of Schistosoma japonicum(Sjcb2) was ampified from the recombinant plasmid pBC SK~+/Sjcb2 by PCR,and cloned into cloning vector pUCm-T,cutting the objective domain by enzymes,then cloned into expression vector pWR450-1 to form the recombinant plasmid.The expression of the recombinant plasmid by SDS-PAGE and Western blotting was analyzed. Results The specific fragment of Sjcb2 was amplified and the recombinant prokaryotic expression plasmid Sjcb2/pWR450-1 was successfully constructed.After being induced by IPTG,the transformed E.coli DH5α expressed an about 86 kDa fusion protein.The results of Western blotting analysis showed that the fusion protein could be recognizd by sera of rabbits with Schistosomiasis japonica. Conclusion The recombinant prokaryotic expression plasmid Sjcb2/pWR450-1 was constructed and the Sjcb2 gene was successfully expressed in E.coli DH5α,which paved the way for further study on the function of Sjcb2.