Construction and expression efficiency of recombinant Bb-EmⅡ/3 vaccine of echinococcus multilocularis
- VernacularTitle:多房棘球绦虫重组Bb-EmⅡ/3疫苗的构建及其表达效率
- Author:
Mei YANG
- Publication Type:Journal Article
- Keywords:
Echinococcus multilocularis;
Recombinant Bb-EmⅡ/3 vaccine;
Construction;
Expression
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct recombinant Bb-EmⅡ/3 vaccine of Echinococcus multilocularis and to investigate the expression efficiency of EmⅡ/3 antigen encoding gene in Escherichia coli BL21(DE3). Methods:EmⅡ/3 antigen gene was amplified by PCR. Then the gene was cloned into Escherichia coli-Bifidobacteria shuttle plasmid pGEX-1?T containing gluathione-S-transferaze(GST) gene to construct pGEX-EmⅡ/3. The recombinant plasmid was electroporated into Bifidobacteria bifidum(Bb) to construct rBb-EmⅡ/3 vaccine. The vaccine was identified with PCR and restriction-endonuclease digestion. The expression of pGEX-EmⅡ/3 was induced with isopropyl-?-D-thiogalactopyranosid(IPTG),and fusion protein EmⅡ/3/GST was examined by SDS-PAGE and Western blot techniques. Results:1 759bp gene of EmⅡ/3 was successfully amplified by PCR and cloned into pGEX-1?T by restriction analysis. rBb-EmⅡ/3 vaccine was successfully constructed according to PCR and restriction-endonuclease digestion.It was demonstrated with SDS-PAGE and Western blot that the fusion protein EmⅡ/3/GST were expressed in E.coli,BL21. Conclusions:rBb-EmⅡ/3 vaccine of Echinococcus multilocularis is successfully constructed. The plasmid pGEX-EmⅡ/3 was highly expressed in E.coli in fused form with GST and this kind of protein shows specific antigenicity according to western blot.
