Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Characterization of the Recombinant Proteins of Porcine ircovirus Type 2 Field Isolate Expressed in the Baculovirus System | J. Vet. Sci. 2002 3 1 19-23 JOURNAL OF Veterinary Science Characterization of the Recombinant Proteins of Porcine Circovirus Type2 Field Isolate Expressed in the Baculovirus System Yuna Kim Jinhyun Kim Kyoungsoo Kang1 and Young S. Lyoo Immnunopathology Lab. College of Veterinary Medicine Konkuk University Seoul 143-701 Korea Dongdoochun Bayer Vet. Hospital Dongdoochun Kyounggi-do Korea ABSTRACT Porcine circovirus PCV type2 was isolated using primary porcine kidney cells from lymph node of piglets with typical PMWS. The presence of the virus was identified by PCR using primers specific to PCV type2. The ORFs 1 and 2 were amplified by PCR using primers corresponding to the target genes of the PCV type 2. Cloned genes were inserted into the baculovirus expression vector and PCV recombinant proteins were expressed using baculovirus expression system. Recombinant protein expression was determined by indirect immunofluorescent assay IFA and immunoblotting using polyclonal antiserum to PCV. ORF1 gene expressed two proteins with approximately 17 kDa and 31 kDa proteins in the baculovirus system. Recombinant protein of the ORF2 was similar to that of the native virus except minor bands with different molecular weight were detected. Recombinant protein expressed in the baculovirus system showed at least two glycosylation sites based on the tunicamycin treatment. Recombinant protein of the ORF2 assembled virus-like particle in recombinant virus infected insect cells. Key words PCV type2 recombinant protein baculovirus system Introduction Post-weaning multisystemic syndrome PMWS has been hot issue in swine industry with controversy of causative agent that may produce this newly recognized pig disease. Among several pathogens detected PCV type2 has been consistently identified from animals with typical PMWS 1 2 10 11 . However non-pathogenic porcine circovirus PCV Corresponding author Dr. Young S. Lyoo Assistant professor Immunopathology Lab. College of Veterinary Medicine .
