Construction and identification of recombinant BHV-1 expressing foot and mouth disease virus VP1 gene

Abstract:

Abstract: [Objective]: In order to construct the recombinant bovine hepervirus-1(BHV-1) which expressed foot and mouth disease virus (FMDV)VP1 gene, we constructed a BHV-1 gE gene transfer vector by inserting the synthetic VP1 gene of FMDV (O/China/99) under the immediate-early promoter of cytomegalovirus. [Methods]: The mixtures of parental virus (BHV-1/gE-/LacZ+ ) DNA and transfer vector was transfected into bovine turbinate cells using calcium phosphate-mediated transfection. Then the propagated viruses were harvested. The recombinant BHV-1 (designated BHV-1/gE-/VP1) was obtained by selection for white virus plaques. [Results]: PCR results showed that VP1 gene was successfully inserted into the genome of BHV-1/gE-. The expression of VP1 in infected cells was proved by indirect immunofluorescence assay and Western blotting. [Conclusion]: The research provided a basis for development of BHV-1 vector vaccines for FMD and other important bovine infectious diseases.