Establishment and detection of tetracycline-inducible stable cell line expressing hepatitis C virus nonstructural protein 5B
WANG Yao-Hui1; DING Huan-Ping2; WANG Yu-Chan1; YUAN Zheng-Hong1,2,3
1. Key Laboratoty of Medical Molecular Virology, Ministry of Education and Health, Shanghai Medical College, Fudan University, Shanghai 200032, China; 2. Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China; 3. Shanghai Public Health Clinical Center Affiliated to Fudan University, Shanghai 201508, China
Abstract：Hepatitis C virus (HCV) infection is a major risk factor of chronic hepatitis, liver cirrhosis and even hepatocellular carcinoma (HCC). However, the detailed mechanism on cancer caused by HCV is still elusive. Nonstructural protein 5B (NS5B) is one of the nonstructural proteins encoded by HCV, and essential for HCV RNA replication because of its RNA-dependent RNA polymerase (RdRp) activity. In addition to participating in viral RNA replication, NS5B was also found to be involved in the pathogenicity of HCV through direct interaction with host proteins. In order to address the role of NS5B in HCV replication and pathogenicity, we constructed a tetracycline-inducible (Tet-on) stable cell line expressing NS5B in HepG2 cells. Protein expression was examined by Western blot using both tag antibody and anti-NS5B antibody induced by doxycycline either at different concentrations or at different indicated time points. Moreover, immunofluorescence assay further confirmed the results. In conclusion, the HepG2 Tet-on NS5B cell line will be a good model to investigate the role of NS5B in HCV replication and pathogenicity.
王耀辉1; 丁焕平2; 王燏婵1; 袁正宏1,2,3. 四环素诱导的丙型肝炎病毒非结构蛋白5B稳定细胞株的构建和检测[J]. 微生物与感染
, 2011, 6(2): 84-89.
WANG Yao-Hui1; DING Huan-Ping2; WANG Yu-Chan1; YUAN Zheng-Hong1,2,3. Establishment and detection of tetracycline-inducible stable cell line expressing hepatitis C virus nonstructural protein 5B. Journal of Microbes and Infections, 2011, 6(2): 84-89.