Clone expression and purification of influenza A virus nucleoprotein gene.
- Author:
Shi-song FANG
1
;
Xiao-wen CHENG
;
Tao LIU
;
Jian-jun LIU
;
Shu-jin ZHAO
;
Li ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Chromatography, Affinity; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Escherichia coli; genetics; Gene Expression; Influenza A virus; genetics; isolation & purification; metabolism; RNA, Viral; genetics; RNA-Binding Proteins; genetics; metabolism; Recombinant Proteins; isolation & purification; metabolism; Reverse Transcriptase Polymerase Chain Reaction; Viral Core Proteins; genetics; metabolism
- From: Chinese Journal of Experimental and Clinical Virology 2005;19(2):165-168
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone the influenza A virus NP gene into expression vector and to purify the target protein, which was used to study the preparation of monoclonal antibody.
METHODSThe RNA of influenza A virus was extracted and primers were designed according the NP gene sequence, then the NP gene of influenza A virus was expressed in E.coli DH5alpha and the NP protein was purified by affinity chromatography.
RESULTSThe recombinant expression vector-pGEX-4T-2-NP was successfully constructed and relatively pure target protein was obtained.
CONCLUSIONThrough reasonably controlling the fermentation time, growth temperature and induction concentration, satisfactory soluble target product was obtained.
