Cloning and expression of NS3 gene of hepatitis C virus in yeast
- VernacularTitle:丙型肝炎病毒NS3基因酵母表达载体构建及表达
- Author:
Ke LI
;
Lin WANG
;
Jun CHENG
- Publication Type:Journal Article
- Keywords:
Hepatitis C-like viruses;
Viral nonstructural proteins;
Yeasts;
Gene expression
- From:
Chinese Journal of Infectious Diseases
1999;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective HCV NS3 protein plays an important role in disease caused by HCV. We investigate the gene expression of HCV NS3 in yeast for future study of the function of the protein. Methods PCR was performed to amplify the gene of HCV NS3 from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector. Thereafter, HCV NS3 gene was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, and recombinant pGBKT7∶NS3 was transformed into yeast AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. Results HCV NS3 gene was successfully cloned into pGBKT7. The results of SDS PAGE and Western blotting assay showed that the molecular weight of the expressed product was about 22000 Da and HCV NS3 protein was existed within yeast cells.Conclusions HCV NS3 was successfully expressed in yeast expression system.
