Designing and construction of a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis

Abstract

Background and Aim : Novel TB vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis .  Materials and Methods: Tb10.4 fragment was amplified by PCR and the products were digested with restriction enzymes. Next, it was cloned into the pcDNA3.1 + plasmid. The ag85a gene and pcDNA3.1 + / tb10.4 plasmid were digested by EcoRI and BamH1 restriction enzymes. Eukaryotic cells were transfected with pcDNA3.1 + / tb10.4 - ag85a plasmid for confirming expression of tb10.4 - ag85a in these cells. Results: Using electrophoresis of PCR products, fragments 297 bp for tb10.4 and 1017 bp for ag85a were observed. Eukaryotic cells transfection with pcDNA3.1 + / tb10.4 - ag85a vector was confirmed with cDNA synthesis and existence of tb10.4-ag85a was confirmed with RT-PCR. Conclusion: In this study, we constructed a DNA vaccine encoding tb10.4-ag85a fusion fragment. It can be used for development of more new DNA vaccines in future studies .