[Transfection and expression of HCV-NS(5)B gene in Huh-7 cells].
OBJECTIVE
Hepatitis C virus (HCV) is the major cause of blood-borne non-A, non-B hepatitis. An RNA-dependent RNA polymerase is encoded by HCV non-structural protein 5B (NS(5)B) gene. A full-length HCV RNA has been transfected into human hepatoma cells, and HCV NS(5)B has been expressed in E.Coli purified. But human hepatocyte line with high expression of NS(5)B has not been established yet.


METHODS
The NS(5)B 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 NS(5)B protein in Huh-7 cells was detected by Western blot analysis.


RESULTS
There were the NS(5)B sequence and the expression of HCV-RNA polymerase in Huh-7 cells transfected NS(5)B plasmids.


CONCLUSIONS
We have established a HCV RNA polymerase expression system in Huh-7 cells that can be used to study the mechanism of HCV replication and to test gene therapy of hepatitis C.
Fang
J
J
Shanghai Institute of Digestive Disease, Shanghai Second Medical University, Shanghai 200001,China.
Edwin
O
O
D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D016133:Polymerase Chain Reaction; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase