Evaluation of the expression pcDNA3.1(+)/ESAT-6 by myoblast cells of BALB/c mice against Bovine Tuberculosis

one of the most common diseases in zoonotic is Bovine tuberculosis. It has been more attended because of prevalence and economic loss. Although, many efforts have been done to produce the vaccine, but the research in this area is going on. Mycobacterium bovis (M. bovis) is the most important pathogen in bovine tuberculosis. The ESAT-6 antigen is considered as a protective immunogenic and also its important candidate for DNA vaccine. BCG vaccine makes a variable immunity period and it is not desirable. Immunization with recombinant plasmid encoding protective proteins is a promising vaccination technique Therefore, the aim of this study was cloning and expression of recombinant protein in the mice myoblast cells of BALB /c as the immunogenicity of candidate. The ESAT-6 antigen from M. bovis was cloned in a eukaryotic expression plasmid pcDNA3.1 (+). The integrity of the constructed plasmid was confirmed by using clone PCR, restriction enzyme mapping and sequencing. Recombinant plasmid pcDNA3.1 (+) /ESAT-6 injected into Myoblast cells. After one week the samples was taken from muscle tissue. The electrophoresis and western blotting confirmed the recombinant protein of ESAT-6 (rESAT-6) in Myoblast cells. Also the structure prediction showed that the recombinant protein antigenicity and hydrophobicity index is high. The current research indicated that rESAT-6 could be used as a research experimental tool in protection assays against bovine tuberculosis.