Expression of the VP2 protein of canine parvovirus 2b using a baculovirus expression system

서지정보

ㆍ발행기관 : 한국예방수의학회(구 한국수의공중보건학회) ㆍ수록지정보 : 예방수의학회지 / 47권 / 3호
ㆍ저자명 : Min-Ji Kim, Seong-In Lim, MiJung Kwon, Min Ji Kim, Eui Hyeon Lim, Bang-hun Hyun, Yoon-Hee Lee

목차

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

영어 초록

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.

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