Study On Expression Of HCV Structural Proteins And The Forming Of HCV-like Particles In Insect Cells

Hepatitis C virus (HCV) infection is a leading cause of chronic hepatitis and hepatocellular carcinoma worldwide. Up to now, there is neither vaccine nor efficient therapy drug available.HCV structural proteins (C, El, E2) which comprise the viron not only play crucial roles in the entrance of HCV into host cells, but also can induce strong immune responses of the host. The study of HCV structural proteins will be helpful for understanding the morphogenesis and the biological characteristics of HCV, as well as for the development of HCV vaccine and diagnostic reagent.This study contains two parts. In part one, different regions of HCV core protein were expressed in E.coli and their immunological characteristics were studied. Part two involved simultaneous expression of all three structural proteins in insect cells. The interactions of expressed proteins were investigated, and the assembly of HCV-like particles were observed.Part one: Expression of hepatitis C virus core protein in E.coli andtheir immunological characteristicsThree cDNA sequences encoding full length of HCV core protein (C191) and two truncated forms (C154 and C69) which lack parts of the carboxyl-terminal were amplified by PCR and inserted into plasmids pQE30 and pTrcHisA. The resulting expression vectors were transformed into 4 strains of E.coli (DH5a, TOP10, BL21, and M15). The produced proteins were detected by SDS-PAGE and Immunoblotting...