Construction and identification of recombinant vaccine Bifidobacterium bifidum pGEX-Sj14-3-3 of Schistosoma japonicum
10.3760/cma.j.issn.1000-4955.2011.04.002
- VernacularTitle:日本血吸虫重组两歧双歧杆菌pGEX-Sj14-3-3疫苗构建及鉴定
- Author:
Ning, ZHANG
;
Wen-gui, LI
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
Bifidobacterium bifidum;
Plasmids;
Vaccines
- From:Chinese Journal of Endemiology
2011;30(4):357-360
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and identify recombinant vaccine Bifwlobacterium bifidum(Bb)pGEX-Sj14-3-3 of Schistosoma japonicum(Sj). Methods Total RNA was extracted from adult Sj, antigen encoding gene Sj14-3-3 was amplified by RT-PCR and cloned into Escherichia coli (E. coli)-Bb shuttle expression vector pGEX-1λT to construct recombinant plasmid pGEX-Sj14-3-3. The recombinant plasmid was transformed into E. coli BL21 (DE3).The plasmid was extracted and identified by using BamH I and EcoR I. Then pGEX-Sjl4-3-3 was electroporated into Bb to construct recombinant Bb (pGEX-Sj14-3-3) vaccine. The extracted plasmid of the recombinant Bb (pGEX-Sj14-3-3) vaccine was identified by PCR, and the size of the products was compared with Sj14-3-3 gene of adult worms.Results Sj14-3-3 of 399 bp in length was amplified by RT-PCR. The products were digested by BamH I and EcoR I , and the fragments length of plasmid pGEX-Sj14-3-3 vector was 4947 bp, and of Sj 14-3-3 gene was 399 bp.The product of 399 bp Sj14-3-3 gene was also amplified by PCR from template of the extracted plasmid of the recombinant Bb(pGEX-Sj14-3-3 ) vaccine. The size of the product obtained was just the same as expected.Conclusion The recombinant Bb(pGEX-Sj14-3-3) vaccine of Sj is successfully constructed.
