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 HCV RNA polymerase has been established.


METHODS
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.


RESULTS
There were NS5B gene fragments and the expression of NS5B protein in Huh-7c cells transfected with pTeT-NS5B or pcDNA-NS5B plasmid.


CONCLUSIONS
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.
Fang
J
J
Shanghai Institute of Digestive Disease, Shanghai Second Medical University, Shanghai 200001, China.
Ouyang
E
E
Wu
C H
CH
Wu
G Y
GY
Qiu
D
D
Xiao
S
S
D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D016174:Hepacivirus / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D014162:Transfection; D014407:Tumor Cells, Cultured; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase