Cloning Of Several Protective Antigen Genes Of Mycobacterium Tuberculosis And Prokaryotic Expression Of MPT-64Gene

Objective: Tuberculosis (TB) is a chronic infectious disease caused by infection ofMycobacterium tuberculosis. Mycobacterium tuberculosis may invade organs of human body,but majority infection of the lungs and be known as tuberculosis. At present, our country is oneof the countries which are affected most seriously by TB in the world, and we have always paidattention to the prevention and cure of TB. At present, our country prevents TB by BCGinjection, but the effect of prevention is not satisfactory. Some researchers have set about tryingto detection tuberculosis antibody by using fusion protein[1]. This experiment selected andcloned of several protective antigen genes of Mycobacterium tuberculosis. To get the MPT-64fusion protein, the MPT-64was transfected into competent cells to express. This experimentwill do matting for TB diagnosis and vaccine development.Method: The researchers the protective antigen genes of Mycobacterium tuberculosis(Ag85a, Ag85b, MPT-63, MPT-64, CFP-10, CFP-21, TB10.4, ESAT-6) was screened, then theprimers of them were designed, and amplificated them by PCR. MPT-64gene was selected fromthem to constructed recombinant plasmid pET-MPT-64-32a, and then expressed it throughprokaryotic cells to obtain the fusion protein.Result:8kinds of protective gene of Mycobacterium tuberculosis (Ag85a, Ag85b, MPT-63,MPT-64, CFP-10, CFP-21, TB10.4, and ESAT-6) were successfully cloned. The recombinantplasmid pET-MPT-64-32a was obtained successfully, and the MPT-64protein was expressed byprokaryotic cells.Conclusion: This experiment got MPT-64fusion protein, and it would be helpful in thediagnosis, treatment of TB and vaccine development.