Transfection and expression of HCV-NS5B gene in Huh-7 cells

Objective To study the mechanism of hepatitis C virus (HCV) replication and to test gene therapy for hepatitis C, a human liver cell line expressed HC V RNA polymerase has been established. Methods NS5B gene has been transfected into Huh-7 cells by lipofectamine. T he results of transfection were confirmed by PCR and Southern blot analysis , and the level of the non-structural protein 5B (NS5B) in Huh-7 cells was detected by Western blot analysis. Results There were NS5B gene fragments and the expression of NS5B protein i n Huh-7c cells transfected with pTeT-NS5B or pcDNA-NS5B plasmid. Conclusions We have established a HCV RNA polymerase expression system in H uh-7 cells which can be further used to analyze the mechanism of HCV replic ation and provide a cell model for gene therapy in vitro.