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KMID : 0380420100340030229
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Journal of Prventive Veterinary Medicine
2010 Volume.34 No. 3 p.229 ~ p.236
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Expression of type 2 porcine circovirus-Like particles of two major genotypic groups in baculovirus recombinants
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Kim Ha-Hyun
Park Geon-Yong
Park Jun-Gyu
Kim Hyun-Jeong
Kwon Hyoung-Jun
Park Su-Jin
Jeong Young-Ju
Kim Chae-Hyun
Myra Hosmillo
Mia Madel Alfajaro
Therese Marie Collantes
Kang Mun-Il
Park Sang-Ik
Cho Kyoung-Oh
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Abstract
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Porcine circovirus type 2 (PCV2) is associated with porcine circovirus diseases (PCVD), of which postweaning
multisystemic wasting syndrome is considered to cause considerable economic losses in pig industry worldwide. As the
virus-like particle (VLP) is a highly effective type of subunit vaccine and has unique advantages in terms of safety and
immunogenicity, this study aimed to develop PCV2-like particles, which matched currently circulating Korean PCV2
and were applicable as vaccines. The ORF2 genes encoding PCV2 capsid protein were amplified from the PCV2
subgroup 1A/B Korean C275 isolate and the subgroup 2E C94 isolate by PCR assay with primer pair specific to PCV2
ORF2 gene, and were cloned into baculovirus transfer vector. Recombinant baculovirus was generated by cotransfection
with the transfer vector and linear baculovirus DNA into the Sf9 cells, and then by plaque purification. Expression of
PCV2 capsid protein was determined by the indirect immunofluorescence and Western blotting assays, and electron
microscopy. By both immunological assays, PCV2 capsid antigen was detected in the Sf9 cells infected with the
recombinant baculoviruses. The formation of empty virus particles, characteristic of VLP, was detected in the lysate of
Sf9 cells infected with the recombinant baculoviruses by negative electron microscopy. From these results, VLPs of two
genogroups of PCV2 were successfully expressed and generated in a baculovirus expression system. It is expected that
the expressed VLPs of two genotypic groups can be used for control of PCV2 infection as good vaccine candidates.
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KEYWORD
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Baculovirus Expression System, Porcine Circovirus Type 2, Vaccine Candidate, Virus-Like Particles
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