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- W2355792011 abstract "Objective To obtain recombinant Staphylococcal enterotoxin B(SEB) by gene engineering and evaluate its biological activity.Methods The full-length of SEB gene amplified from DNA of S6 strain of Staphylococcus aureus by high-fidelity PCR was cloned into prokaryotic expression vector pET22b~+,and then the vector was transformed into BL21(DE3) to construct a prokaryotic expression system.The sequence of SEB gene was confirmed by DNA sequencing and the recombinant SEB(rSEB) was induced by 1 mmol/L IPTG.The expressed product was identified by SDS-PAGE and the expressed rSEB was purified by HiTrap~(TM) chelating HP.The antigenicity of rSEB was tested by ELISA and the toxicity of rSEB was tested by using a lipopolysaccharide(LPS) and actinomycin D sensitized mouse model.Results The nucleotide sequence of the cloned SEB gene was the same as that of reported in Genebank.The relative molecular weight shown on SDS-PAGE profile was consistent with expected value.The purified rSEB was obtained.Positive immune reaction was occurred between the rSEB and the monoclonal antibody of anti-wild type SEB.The lethality assay in mice demonstrated that toxicity of rSEB was the same as that of wild type SEB.Conclusion The gene of SEB is successfully cloned into plasmid pET22b and expressed in E.Coli,which would provide a basis for analyzing the to-(xicity)-related active sites in SEB molecule and further studies on the pathogenesis of SEB." @default.
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- W2355792011 date "2006-01-01" @default.
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- W2355792011 title "Cloning and expression of Staphylococcal enterotoxin Bgene" @default.
- W2355792011 hasPublicationYear "2006" @default.
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