Cloning and Expression of Hepatitis B Surface Antigen

Background and Aims: Hepatitis B virus (HBV) is a major cause of both acute and chronic liver disease. It is estimated that there are 350 million carriers of the virus in the world, and a high proportion will develop serious liver disease, including hepatocellular carcinoma. The aim of this study was cloning and expression hepatitis B surface antigen (HBsAg) gene to design a DNA vaccine. Methods: In this study, we amplified the HBsAg gene from Iranian patients. The gene was cloned in pGEMEX-1 expression vector and recombinant plasmid was transformed in to JM109 E. coli strain and induced by IPTG. Results: We amplified, cloned and expressed hepatitis B virus surface antigen successfully and expressed protein was serologically assayed using gel diffusion and western blot analysis. Gene was sequenced and submitted to GenBank. Condusions: The cloned HBsAg gene is ready for using in experimental DNA vaccine animal study. There are some mutations on this recombinant protein (T45D, Y206C and S207R) which will affect on folding and function of recombinant protein.