Study of Cell Culture System Transfected with Full-length cDNA Clone of Hepatitis C Virus*

A Vero-E6 culture system was developed to assay Hepatitis C virus (HCV) replication by plasmid (pHCV) transfection，which contains a T7 promoter at the 5’end, full-length cDNA of the HCV genome and a T7 terminator. To facilitate high-level transcription of HCV RNA, Vero-E6 cells were transfected with pHCV and then infected with recombinant vaccinia virus (vTF7-3) containing the T7 RNA polymerase gene. This transfection-based cell culture system produced high levels of HCV genome (107-108 copies/mL) detectable by real-time quantitative PCR and the production of HCV RNA transcripts was confir med by RT-PCR. Western blot analysis revealed that HCV structural and non-structural proteins were correctly processed. The transfected Vero-E6 cells assembled 40-50nm virus-like particles were analysed with transmission electron microscope. This model can be utilized for studying mechanisms of HCV replication, preparation of HCV vaccine and to test potential antiviral drugs.