Gene cloning and expression of Japanese encephalitis virus E protein in Escherichia coli
- VernacularTitle:乙型脑炎病毒E蛋白基因的克隆及其在大肠埃希菌中的表达
- Author:
Yushui WU
;
Junping REN
;
Wei ZHANG
- Publication Type:Journal Article
- Keywords:
Encephalitis virus, Japanese;
E protein;
Gene expression;
Escherichia coli
- From:
Chinese Journal of Infectious Diseases
2000;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express Japanese encephalitis virus (JEV) E protein in Escherichia coli. Methods The gene coding for JEV E protein was amplified by using RT-PCR, and cloned into plasmid pET-28a. The constructed plasmid was transformed into E.coli BL21 (DE3). Expression of E protein by the transformants was induced by IPTG and analyzed by SDS-PAGE and Western blot. Results Five nucleotide changes leading to amino substitutions were identify in the E protein gene of our laboratory strain of JEV when compared to a published gene sequence of JEV E protein. The yield of expressed JEV E protein with relative molecular mass approximately 52?10 3 was 35% of total bacterial proteins. Conclusions JEV E protein was expressed successfully in E.coli, which should be useful for the production of diagnostic reagents and the analysis of gene structure/function of the E protein.
