Construction of a vector of epitope vaccine GILE oral Lactococcus-lactis for Echinococcus multilocularis with M-cell targeting
Objective To construct a vector of epitope vaccine GILE oral Lactococcus lactis for Echinococcus multilocularis with M-cell targeting.Methods The SAM-GILE sequence was designed and synthesized by adding the SAM gene to the existing multi-epitope vaccine GILE to construct Lactococcus lactis expression plasmid pNZ8148-SAM-GILE.The expression system for Lactococcus lactis was established by electroporating into Lacto-bacillus lactis NZ9000.Double experiments of enzyme digestion and gene sequencing were used to verify whether Lactococcus lactis expression plasmid pNZ8148-SAM-GILE was constructed successfully.PCR were used to verify whether Lactococcus lactis expression plasmid pNZ8148-SAM-GILE was electroporated into Lactococcus lactis NZ9000 successfully.Western Blot were used to detect whether the recombinant protein SAM-GILE was ex-pressed.The surface display of SAM-GILE was identified via whole-cell ELISA.The M-cell targeting of LL-plSAM-GILE was identified by immunofluorescence.Results Double experiments of enzyme digestion and gene se-quence showed the successful construction of Lactococcus lactis expression plasmid pNZ8148-SAM-GILE.PCR showed that pNZ8148-SAM-GILE was successfully electroporated into Lactococcus lactis NZ9000.Western Blot showed that the recombinant protein was expressed successfully at approximately 45 KD induced by Nisin.Whole-cell ELISA detection showed that SAM-GILE could be displayed on the surface of LL-plSAM-GILE.Immunofluo-rescence verified that the recombinant antigen was highly consistent with M cell,indicating that the recombinant Lactococcus lactis vaccine LL-plSAM-GILE had M-cell targeting.Conclusion A vector of epitope vaccine GILE oral Lactococcus lactis for Echinococcus multilocularis with M-cell targeting was successfully constructed.
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