Construction and expression efficiency of recombinant Bb-Em14-3-3 vaccine of Echinococcus multilocularis
- VernacularTitle:多房棘球绦虫重组Bb-Em14-3-3疫苗的构建及其表达效率
- Author:
Mei YANG
;
Wengui LI
;
Youming ZHU
- Publication Type:Journal Article
- Keywords:
echinococcus multilocularis;
recombinant Bb-Em14-3-3 vaccine;
construction;
expression
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
1981;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct recombinant Bb-Em14-3-3 vaccine of Echinococcus multilocularis and to investigate expression efficiency of Em14-3-3 antigen encoding gene in Escherichia coli BL21(DE3).Methods Em14-3-3 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-Em14-3-3.The recombinant plasmid was electroporated into Bifidobacteria bifidum(Bb) to construct rBb-Em14-3-3 vaccine.The vaccine was identified with PCR and restriction-endonuclease digestion.The expression of pGEX-Em14-3-3 was induced with isopropyl-?-D-thiogalactopyranosid(IPTG) and fusion protein Em14-3-3/GST was examined by SDS-PAGE and Western blot techniques.Results 530 bp gene of Em14-3-3 was successfully amplified by PCR and cloned into pGEX-1? T by restriction analysis.rBb-Em14-3-3 vaccine was successfully constructed by PCR and restriction-endonuclease digestion.It was demonstrated with SDS-PAGE and Western blot that the fusion protein Em14-3-3/GST was expressed in E.coli,BL21.The expression efficiency is 23%.Conclusion rBb-Em14-3-3 vaccine of Echinococcus multilocularis was successfully constructed.The plasmid pGEX-Em14-3-3 was highly expressed in E.coli in fused form with GST and this kind of protein shows specific antigenicity.
