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

Chin Med J (Engl). 2000 Jul;113(7):602-5.

Abstract

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.

MeSH terms

  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / metabolism*
  • Carcinoma, Hepatocellular / virology
  • Hepacivirus / physiology
  • Humans
  • Liver Neoplasms / genetics
  • Liver Neoplasms / metabolism*
  • Liver Neoplasms / virology
  • Transfection
  • Tumor Cells, Cultured
  • Viral Nonstructural Proteins / biosynthesis*
  • Viral Nonstructural Proteins / genetics*
  • Virus Replication

Substances

  • Viral Nonstructural Proteins
  • NS-5 protein, hepatitis C virus