Cholera Toxin B Subunit (ctxb) Expression In Shigella

Cholera and dysentery are worldwide epidmic diseases. Presently,the vaccine against cholera and dysentery respectively has existed. Butthere are no the vaccines against the two contagious diseases in the sametime. In our experiment,we construct a multivalent candidate vaccine whichprevent the two diseases.Firstly, we obtain the Hg resistance gene from the strain DH5 a (pMT999)by whole-strain PCR. Then Hg resistance gene was cloned into T-vector andthe recombinant plasmid named pMDHg04.The plasmids pMDHg04, pMGL102, pBR322were digested by the appropriate enzymes and the segments (Hg resistancegene, ctxB and the duplicon of the plasmid pBR322)were recovered, ligatedand transformed. In the end, a new expression vector pBRC09 wasconstructed, in which the Hg resistance gene as a screening tag.Secondly, by electroporation the expression vector pBRC09 wastransferred into the strain 2457TDI1.2457TDI1 is a bivalent Shigella vaccinewith host-vector balanced-lethal system. The experiments such as ELISA andWestern blot demonstrated that CtxB could be expressed high in the newrecombinant strain, which was named 2457TC.Thirdly, the rabbit was immunized with 2457TC. The result indicated that2457TC is a hopeful trivalent candidate vaccine.