Construction of the MTB41 DNA Vaccine of Mycobacterium tuberculosis and Its Cloning,Expression in Prokaryotic Expression Vector

The purpose of this study is to construct the MTB41 DNA vaccine of Mycobacterium tuberculosis and clone it into prokaryotic expression vector to express. The gene encoding MTB41 was amplified from M.tuberculosis H37Rv chromosomal DNA by using PCR technique. PCR product was cloned seperately into the pJW4303 and pET22b, then transformed into E.coli DH5α strain, plasmid DNA was extracted and digested with enzymes.Plasmids containing the right insertion were sequenced to confirm their identity and retransformed the recombinant MTB41+pET22b into E.coli. BL21(DE3)PLysS strain. Bacterial lysates prepared from IPTG induced cultures were loaded directly onto SDS-PAGE. Upon IPTG induction, the recombinant MTB41+pET22b produced a protein with an apparent MW of 41 ku.In conclusion, we have successfully constructed MTB41 DNA vaccine and its protein has been expressed. It will establish the detecting of immunity effectiveness and preparation of the antigen and antibody of MTB41 protein on a large scale.