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.

ABSTRACT
 서 론
 재료 및 방법
  1. 바이러스 및 세포
  2. PCV2 ORF2 유전자의 증폭 및 클로닝
  3. 재조합 baculovirus 제작
  4. Sf9 세포에서 PCV2 VLP 발현
  5. 간접면역형광항체법을 이용한 PCV2 VLP 발현 확인
  6. Western blotting 기법을 이용한 PCV2 VLP 발현 확인
  7. PCV2 VLPs의 정제 및 투과 전자 현미경 관찰
 결 과
  1. 재조합 baculovirus의 ORF2 아미노산 서열 확인
  2. 간접면역형광항체법을 이용한 재조합 baculovirus가감염된 Sf9 세포에서 캡시드 단백질의 발현 확인
  3. Western blotting 기법을 이용한 재조합 baculovirus가감염된 Sf9 세포에서 캡시드 단백질의 발현 확인
  4. 투과전자현미경을 이용한 VLPs 형성 확인
 고 찰
 참고문헌

• 김하현(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Ha-Hyun Kim (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 박건용(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Geon-Yong Bak (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 박준규(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Jun-Gyu Park (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 김현정(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Hyun-Jeong Kim (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 권형준(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Hyoung-Jun Kwon (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 박수진(한국생명공학연구원) | Su-Jin Park (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology)
• 정영주(㈜고려비엔피) | Young-Ju Jeong (KBNP, Inc.)
• 김채현(㈜고려비엔피) | Chae-Hyeon Kim (KBNP, Inc.)
• Myra Hosmillo(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀)
• Mia Madel Alfajaro(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀)
• Therese Marie Collantes(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀)
• 강문일(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Mun-Il Kang (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University)
• 박상익(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Sang-Ik Park (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University) Correspondence
• 조경오(전남대학교 수의과대학 바이오치료 산업인력 양성사업팀) | Kyoung-Oh Cho (Bio-therapy Human Resources Center, College of Veterinary Medicine, Chonnam National University) Correspondence