Construction of eukaryotic expression vector expressing hepatitis B virus HBsAg and EGFP fusion protein and establishment of stable transfected Chang Liver cell line
10.3760/cma.j.issn.1003-9279.2008.03.024
- VernacularTitle:乙肝病毒HBsAg-EGFP融合蛋白真核表达载体构建及稳定转染Chang Liver细胞系的建立
- Author:
Jin-Song MU
1
;
Hui-Fen WANG
;
Jiang-Hua WANG
;
Xiao-Ben PAN
;
Lu ZHANG
;
Lai WEI
Author Information
1. 解放军军医进修学院
- Keywords:
Hepatitis B virus;
Hepatitis B surface antigesn;
Transfection;
Cell line
- From:
Chinese Journal of Experimental and Clinical Virology
2008;22(3):228-230
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a eukaryotic expression vector for expressing hepatitis B virus (HBV) recombinant HBsAg-EGFP fusion protein and obtain a stable transfocted Chang Liver cell line. Methods The coding region of HBsAg gene of HBV was amplified by PCR and was digested by BamH I/EcoR I. This fragment was inserted into pEGFPN1 with T4 ligase and transformed E-coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfocted into Chang Liver cell by Lipofectamine 2000 cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level HBsAg-EGFP fusion protein was obtained. Results The eukaryotic expression vector named pEGFPN1-HBsAg was successfully constructed and the stable transfected Chang Liver celI line could express pEGFPN1-HBsAg fusion protein was obtained. Conclusion The stable transfected Chang Liver cell line could express pEGFPN1-HBsAg fusion protein, could be used to screen the proteins differentially expressed in HBsAg expression Chang Liver cells, which brought some new clues for studying the potential molecular mechanism of HBsAg protein.
