The invention belongs to the technical field of virology and gene engineering, and particularly relates to construction, vaccine preparation and application of recombinant pseudotype baculovirus expressing rabies virus main immunogenic glycoprotein (RVG). The recombinant pseudotype baculovirus obtained in the invention contains two expression cassettes of not only an insect cell expression cassette in which RVG expression is driven by a PPH promoter and RVG is displayed on a virion envelop surface, but also a mammal cell expression cassette in which high-efficient expression of RVG in mammal cells is driven by a CMV promoter. A rabies vaccine prepared by the recombinant baculovirus has dual properties of subunit vaccines and live viral vector vaccines. After mouse immunization, the rabies vaccine can stimulate the mouse body to generate high-level rabies virus specific ELISA antibodies, neutralizing antibodies, and IFN-gamma, and has potential vaccine application value. The recombinant baculovirus BV-RVG/RVG is preserved in China Center for Type Culture Collection (CCTCC) with a preservation number of CCTCC NO: V201022. The invention also comprises preparation and application of the recombinant baculovirus vaccine.