Compavison of JEV E-Hsp70 and E-Bindingdomain Fusion Protein on Immune Responses in Mice

Further investigation of this interesting adjuvant-free carrier effect is necessary to ascertain whether peptide-binding portion alone can replace the whole M.tuberculosis Hsp70 acting as carrier. In our study, we selected a portion of E protein with higher index of antigenic determinants dependent on analysis of computer software and con-structed two chimeric vectors of pPICZα-E-Hsp70 and pPICZα-E-BD. Vectors were transformed into yeast X-33 by electroporation. Expression of fusion protein in yeast was induced by the addition of methanol every 24 hours and analysed by SDS-PAGE and Western blotting. We produced E-Hsp70 fusion protein at a yield of 33 mg per litter of culture and E-Bindingdomain fusion protein at a yield of 97 mg per litter of culture in methylotrophic yeast Pichia pastoris.with specific antigenicitys. To compare the immune response induced by E-Hsp70 fusion protein with that induced by E-bindingdomain, mice were immunized i.p. on day 0 and day 21. Mice immunized with E-bindingdomain had highe