The expression and purification of hepatitis C virus core protein in prokaryotic cells

Hepatitis C virus (HCV) is an enveloped, positive stranded RNA virus. The World Health Organization estimates that there are more than 170 million HCV infected individuals worldwide. Aside from the high morbidity and mortality of the HCV infection, the treatment costs can be a burden on health economics. Core protein is the structural unit of the viral capsid. It also seems to be important for the viral pathogenesis by interfering with various cellular processes. In this study, the first 178 amino acids coding sequence of the core protein was amplified using polymerase chain reaction (PCR) and cloned into the pGEM-T Easy TA cloning vector. Following digestion with BamHI and HindIII, the insert was subcloned into the pQE-30 vector. The protein expression was carried out in the E. coli M15 (prep4) using IPTG as inducer. The bacterium was pelleted with centrifugation and resuspended in 8M urea. The purification of the recombinant protein was made with affinity chromatography (Ni- NTA). The detection of the expressed and purified protein was made with western blotting using chemiluminescent substrate. The expected 21kDA protein band was detected after the expression and purification. In this study, the core protein was expressed in E. coli. The purified protein can be used in pathogenesis studies and in serologic tests as antigen. The cloned vector can be used as a positive control in PCR experiments.