Prokaryotic expression of P gene from rabies virus and application of the indirect ELISA assay in the detection of its gene products
10.3969/j.issn.1002-2694.2010.02.017
- VernacularTitle:狂犬病病毒P基因的原核表达及间接ELISA方法的应用
- Author:
Gang ZHAO
;
Gang LI
;
Tieqiao CHEN
;
Xiaojuan FAN
- Publication Type:Journal Article
- Keywords:
rabies virus;
P gene;
P protein;
prokaryotic expression;
indirect ELISA
- From:
Chinese Journal of Zoonoses
2010;(2):163-167
- CountryChina
- Language:Chinese
-
Abstract:
The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank. The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digestion by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a(+)to obtain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell with IPTG induction. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, the indirect ELISA assay for the detection of rabies virus antibodies in canine serum was applied after management of the optional working condition.
