Inducible expression of non-structural protein 3 of hepatitis C virus in E. coli.
- Author:
Jun CHENG
1
;
Yanwei ZHONG
;
Yan LIU
;
Jing DONG
;
Jizhen YANG
;
Shouchun YANG
Author Information
- Publication Type:Journal Article
- MeSH: Escherichia coli; genetics; Gene Expression; Hepatitis C Antibodies; biosynthesis; genetics; Polymerase Chain Reaction; Recombinant Proteins; biosynthesis; genetics; Viral Nonstructural Proteins; biosynthesis; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2002;16(1):85-87
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo express recombinant non-structural protein 3 of hepatitis C virus (HCV) in E. coli.
METHODSThe non-structural 3 (NS3) region DNA fragment of HCV was amplified by polymerase chain reaction (PCR) and inserted into inducible proeukaryotic expressive vector pET 30C(+)at Bam H1/EcoR1 sites. The competent BL21 (DE3) E.coli was transformed, and then cultured and induced with IPTG. The expressed HCV NS3 protein was confirmed with ELISA and dot blot hybridization using HCV NS3-specific single chain Fv (ScFv) antibody.
RESULTS1 893 bp DNA fragment of HCV NS3 coding region was amplified by PCR technique. HCV NS3 expressive vector pET-NS3 was constructed. After transformation with pET-NS3 and induction with IPTG, recombinant HCV NS3 protein was expressed and confirmed by specific ELISA and dot blot hybridization.
CONCLUSIONSThe recombinant HCV NS3 can be expressed in E. coli.
