Construction of the tetracycline regulating expression system of HBV-S gene and its expression in eukaryotic cells
- VernacularTitle:乙型肝炎病毒S基因四环素调控体系的构建及其在真核细胞中的表达
- Author:
Yu LEI
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
Tetracycline;
Regulating expression
- From:
Journal of Chongqing Medical University
2007;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct tetracycline regulating expression system of HBV-S gene,and study its regulating expression of HBVAg by tetracycline in HepG 2 cells. Methods:Insert HBs gene into the multiple cloning site of pcDNA4,which contain tetracycline operator in the promoter,and the constructed vector with pcDNA6 was co-transferred into HepG2 by means of lipofectamine,and use 400?g/ml Zeocin and 10?g/ml Blasticidin to kill the untransfected host cell line. After 20 days’select,the cell line which express HBsAg stablely after induced by tetracycline was generated. Then the HBsAg expressing was induced by varying concentration of tetracycline(0.5?g/ml,1.0?g/ml,2.0?g/ml,3.0?g/ml).Amount of HBsAg was detected by means of MEIA (Microparticle Enzyme Immunoassay) to determine the appropriate concentration of tetracycline. Result:HBsAg had been expressed at low quantity without tetracycline in transfected HepG2. However the S/N of HBsAg were high but not different after induced by varying concentration of tetracycline. Conclusion:Tetracycline regulating expression system of HBV-S gene was constructed successfully and HBsAg was highly expressed in the vector induced by tetracycline.
