Transfection and expression of HCV-NS5B gene in Huh-7 cells
Fang, J.; Ouyang, E.; Wu, C.H.; Wu, G.Y.; Qiu, D.; Xiao, S.
Chinese Medical Journal 113(7): 602-605
2000
ISSN/ISBN: 0366-6999 PMID: 11776027 Document Number: 514225
To study the mechanism of hepatitis C virus (HCV) replication and to test gene therapy for hepatitis C, a human liver cell line expressed HCV RNA polymerase has been established. NS5B gene has been transfected into Huh-7 cells by lipofectamine. The 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. There were NS5B gene fragments and the expression of NS5B protein in Huh-7c cells transfected with pTeT-NS5B or pcDNA-NS5B plasmid. We have established a HCV RNA polymerase expression system in Huh-7 cells which can be further used to analyze the mechanism of HCV replication and provide a cell model for gene therapy in vitro.