Eukaryotic expression of the core gene of hepatitis C virus genotype 1a

Abstract

Background : Worldwide, hepatitis C virus (HCV) infection is a serious public health disease unlike hepatitis A and B, there is currently no vaccine against HCV available. Thus, extensive studies are under way to design new and effective treatments against HCV. Core protein is a component of HCV particle which is the first antigen recognized by the immune system.beside protective properties of core protein, anti –core antibodies can be used to monitor the disease progress. The purpose of the present study was to isolate and clone the core (C) gene from HCV genotype 1a in an attempt to construct a recombinant vector and subsequently evaluate its expression in a cell culture system. Methods: RNA genome of HCV genotype 1a was extracted from the blood of an infected patient. Complementary DNA (cDNA) was synthesized. HCV 1a core gene was amplified by PCR using specific primers and it was cloned into a eukaryotic expression vector. Huh7.5 cells were transfected by the designed recombinant vector and the cellular expression of the core gene was confirmed by RT-PCR. Results : Recombinant pcDNA3.1 (+) vector containing the HCV core gene with approximate size of 576bp was successfully designed. RT-PCR was used to confirm the expression of core antigen in an Huh7.5 cell line. Conclusion : The results showed that the core gene was successfully isolated from HCV genotype 1a and was cloned into the eukaryotic expression vector. This recombinant vector effectively replicated in Huh7.5 cell line. and its protective and therapeutic effects can be examined in further investigations.