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Transfection and expression of HCV-NS5B gene in Huh-7 cells |
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| Authors: | FANG Jingyuan Edwin Ouyang Catheine HWu |
| Institution: | Shanghai Institute of Digestive Disease, Shanghai Second Medical University, Shanghai 200001, China;Division of GI, Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA;Division of GI, Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA;Division of GI, Department of Medicine, University of Connecticut Health Center, Farmington, CT 06030, USA;Shanghai Institute of Digestive Disease, Shanghai Second Medical University, Shanghai 200001, China;Shanghai Institute of Digestive Disease, Shanghai Second Medical University, Shanghai 200001, China |
| 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. |
| Keywords: | HCV-NS5B Huh-7 cells transfection PCR Western blotting |
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