[Clone expression and purification of influenza A virus nucleoprotein gene].
OBJECTIVE
To 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.


METHODS
The 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.


RESULTS
The recombinant expression vector-pGEX-4T-2-NP was successfully constructed and relatively pure target protein was obtained.


CONCLUSIONS
Through reasonably controlling the fermentation time, growth temperature and induction concentration, satisfactory soluble target product was obtained.
Fang
Shi-song
SS
Department of Food and Bioengineering, South China University of Technology, Guangzhou 510642, China.
Cheng
Xiao-wen
XW
Liu
Tao
T
Liu
Jian-jun
JJ
Zhao
Shu-jin
SJ
Zhou
Li
L
D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D009980:Influenza A virus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D019590:Nucleocapsid Proteins; D012367:RNA, Viral / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism
C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins