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Construction of a dual-promoter expression plasmid delivered by Salmonella choleraesuis C500 / 生物工程学报
Chinese Journal of Biotechnology ; (12): 341-347, 2009.
Article Dans Chinois | WPRIM | ID: wpr-302815
ABSTRACT
Salmonella choleraesuis C500 strain is an attenuated vaccine preventing piglet from paratyphoid and can also be used as a live vector of other DNA vaccines. Through mucosal immunization, immune response to specific antigens carried by it can be induced. To enhance the immune efficiency of DNA vaccine it carried, promoter Ptrc was inserted into the down stream of the human cytomegalovirus (CMV) immediate early promoter of eukaryotic expression plasmid pEGFP-C1. Then transcription terminator rrnbT1T2 was inserted into down stream of the multiple clone sites of pEGFP-C1, and the dual-promoter expression vector pEGFPPtrcR was constructed. Using 1xTSS method, we transformed the recombinant plasmid into C500, and obtained C500/pEGFPPtrcR. We used SDS-PAGE and Western blotting to detect the expression of report gene EGFP. Strong green fluorescence was observed under fluorescent microscope. The stable passages of this recombinant bacterium were at least 20 generations in vitro. Using liposome we transfected plasmid pEGFPPtrcR into Vero cell. After 24 h, green fluorescent was observed, showing the expression of EGFP in nuclei and endochylema. The construction of dual-promoter expression vector pEGFPPtrcR was successful. The foreign gene was expressed in Salmonella strain C500 and somatocytes, resulting in increased antigen expression. This research provides a foundation for the research of new DNA vaccines which use Salmonella C500 as carrier.
Sujets)
Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Plasmides / Salmonella arizonae / Suidae / Protéines recombinantes / Régions promotrices (génétique) / Vaccins à ADN / Vaccins antisalmonella / Protéines à fluorescence verte / Allergie et immunologie / Vecteurs génétiques Limites du sujet: Animaux langue: Chinois Texte intégral: Chinese Journal of Biotechnology Année: 2009 Type: Article

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Texte intégral: Disponible Indice: WPRIM (Pacifique occidental) Sujet Principal: Plasmides / Salmonella arizonae / Suidae / Protéines recombinantes / Régions promotrices (génétique) / Vaccins à ADN / Vaccins antisalmonella / Protéines à fluorescence verte / Allergie et immunologie / Vecteurs génétiques Limites du sujet: Animaux langue: Chinois Texte intégral: Chinese Journal of Biotechnology Année: 2009 Type: Article