Expression of recombinant Hepatitis C virus (HCV) ore, E1 and E2 proteins by the baculovirus expression vector system

Hepatitis C virus (HCV) infection is definitely a global severe health problem. Neither a vaccine to prevent the infection nor an effective treatment for all HCV genotypes is available at the present time. The prophylactic HCV vaccine researches are recently focusing on eliciting antibody responses to three highly immunogenic structural proteins containing core protein (C) and envelope glycoproteins E1 and E2. The synthesis of the structural proteins-containing virus-like particles (VLPs) may provide us a useful tool to clarify the structural requirements for the assembly of HCV particles. HCV like particles also draw attention for their potential role in HCV vaccine development. In this study, HCV genotype-1a genomic RNA from the serum sample of a chronically-infected patient subjected to reverse transcription polymerase chain reaction (RT-PCR) and cloning into baculovirus pFastBacHTB vector. After vector's restriction analysis and sequencing for verification, the Bac-to-Bac system successfully generated the CE1E2 recombinant bacmid, which then transfected into Sf9 insect cells to produce the recombinant baculovirus. Finally, analysis through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting demonstrated the expression of recombinant CE1E2 protein. The described system represents an efficient means of simultaneous HCV structural proteins expression which may potentially be used for vaccine development and/or diagnostic purposes.