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.