Canine parvovirus-2 (CPV-2) has been reported worldwide as a major pathogen associated with acute hemorrhagic enteritis. The disease is a major infectious cause of death, particularly in young dogs. The earliest type of CPV-2 was replaced with three main subspecies, CPV-2a, CPV-2b, and CPV-2c, within a few years. Vaccination is carried out regularly, but the emergence of antigenic variants and the influence of maternal antibodies have limited the efficacy of commercial vaccines. New vaccines, such as the subunit vaccine, have been developed for alternative, safe, and effective vaccination. The baculovirus expression vector system (BEVS) is an excellent eukaryotic expression system with a high-level expression of foreign proteins and the ability of post-translational modification. Therefore, it is used widely to produce recombinant protein and subunit vaccines. In this study, the VP2 protein of CPV-2b cloned in the gateway vector system was generated using a baculovirus expression system in Spodoptera frugiperda (SF9) insect cells. Hemagglutination assay (HA) titers (24) were obtained, and the expression was detected in 6-His tagged VP2 and monoclonal antibody (mAb) against CPV-2 by western blotting. The VP2 protein of CPV-2b expressed in this study may provide a basis for a clinical diagnosis and vaccination applications for CPV-2.

INTRODUCTION
MATERIALS AND METHODS
    Cells and Virus
    Construction of recombinant transfer vector
    Transfection and production of recombinant baculovirusin SF9 cell
    DAB staining
    SDS-PAGE and Western blotting
    Hemagglutination assay (HA)
RESULTS
    Identification of recombinant bacteria containing positivebacmids
    Production of recombinant baculovirus
    Expression and characterization of VP2 protein againstCPV-2b
DISCUSSION
ACKNOWLEDGEMENTS

• Min-Ji Kim(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• Seong-In Lim(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• MiJung Kwon(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• Min Ji Kim(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• Eui Hyeon Lim(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• Bang-hun Hyun(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea)
• Yoon-Hee Lee(Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbuk-do, 39660, Korea) Corresponding Author