Construction and identification of recombinant Bb-Eg95 vaccine of Echinococcus granulosus
10.3760/cma.j.issn.1000-4955.2009.02.029
- VernacularTitle:细粒棘球绦虫重组Bb-Eg95疫苗的构建及鉴定
- Author:
Bi-ying, ZHOU
;
Ya-tang, CHEN
;
Wen-gui, LI
;
Mei, YANG
- Publication Type:Journal Article
- Keywords:
Echinococcus granulosus;
Vaccines,synthetic;
DNA,recombinant
- From:Chinese Journal of Endemiology
2009;28(2):218-221
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and identify recombinant Bifutobacteria (rBb)-Eg95 vaccine of Echinococcus granulosus (Eg). Methods The total RNA was extracted from hydatid cyst protoscoleces shattered by ultrasound, Eg95 antigen encoding gene was obtained by reverse transcription-polymerase chain reaction(RT-PCR) from the template of total RNA using the primer designed according to the DNA sequence of Eg95, the gene was cloned into Escherichia coli-Bifutobacteria(E.coli-Bb) shuttle plasmid pGEX-1λT and transformed into E.coli BL2 (DE3) competent cell to construct recombinant plasmid pGEX-Eg95 using BamH Ⅰ and EcoR Ⅰ, the recombinant plasmid was identified by restriction endonuclease digestion, then was electroporated into Bb to construct rBb-Eg95 vaccine, the vaccine was identified by PCR. Results Four hundred and seventy-one bp Eg95 gene was amplified by RT-PCR, the products of restriction endonuclease digestion were the same as expected(471 bp Eg95 gene and 4947 bp pGEX-1λT), 471 bp Eg95 gene fragment was amplified by PCR from the template of pGEX-Eg95 extracted from rBb vaccine. Conclusion rBb-Eg95 vaccine of Eg is successfully constructed, which lays the theoretical foundation for exploitation and utilization of this vaccine.
