Abstract:

To develop genetic engineering living-vector vaccine of Bovine viral diarrhea-mucosal disease virus (BVDV)， E₂ gene of BVDV Changchun184 strain was selected and the complete sequence was analyzed，then the epitopes of E₂ protein were predicted by bioinformatics software.Six pairs of primers were designed according to E₂ gene sequence，and the epitopes coding regions (1-297，1-345，1-374，45-297，45-345，45-374) of E₂ gene were amplified by PCR and cloned into pMV361 vector.PCR，restriction endonuclease digestion and sequence analysis proved that the 6 fragments were inserted into the expected position.It was indicated that the six prokaryotic expression plasmids were constructed.The positive recombinant plasmids were electro-transformed into BCG for expression at 45 ℃.The SDS-PAGE and Western blot results indicated that the recombinant proteins could react with polyclonal antibody against BVDV.Our study provides a basis for the further studies on genetic engineering living-vector vaccine of BVDV.