Detection of HBsAg gene of hepatitis B virus by polymerase chain reaction

2. ABSTRACT In the present study, Hepatitis B (HBV) surface antigen (HBsAg) was amplified by polymerase chain reaction (PCR) and cloned into an expression vector. Firstly, HBV DNA was extracted from serum a patient with chronic HBV. Then, HBsAg gene was placed into pcDNA3.1/V5 His_TOPO vector. The resulting recombinant plasmid was used for transformation of E. Coli cells. The presence of HBsAg gene in the vector was confirmed by PCR- Screening assay. Furthermore, Restriction enzyme cutting assay was performed fot the same purpose. So HBsAg gene of HBV was cloned into an expression vector. The recombinant vector will be used for further studies such as DNA immunization and ELISA development in the future. Key words: Hepatitis B virus, polymerase chain reaction, cloning.