[Inducible expression of non-structural protein 3 of hepatitis C virus in E. coli].
BACKGROUND
To express recombinant non-structural protein 3 of hepatitis C virus (HCV) in E. coli.


METHODS
The 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.


RESULTS
1 893 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.


CONCLUSIONS
The recombinant HCV NS3 can be expressed in E. coli.
Cheng
Jun
J
Gene Therapy Research Center, Institute of Infectious Diseases,302 nd Hospital of PLA, Beijing 100039, China.
Zhong
Yanwei
Y
Liu
Yan
Y
Dong
Jing
J
Yang
Jizhen
J
Yang
Shouchun
S
D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D018937:Hepatitis C Antibodies / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D018937:Hepatitis C Antibodies; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases