한국동물위생학회지 (Korean Journal of Veterinary Service)

  • 제32권1호
  • /
  • Pages.19-25
  • /
  • 2009
  • /
  • 3022-7372(pISSN)
  • /
  • 2287-7630(eISSN)
한국동물위생학회 (The Korean Society of Veterinary Service)
권호 표지

돼지생식기호흡기증후군 바이러스의 ORF6 유전자 발현

Expression of ORF6 gene of porcine reproductive and respiratory syndrome (PRRS) virus

  • 배수정 (충북대학교 수의과대학 및 동물의학연구소) ;
  • 김진원 (충북대학교 수의과대학 및 동물의학연구소) ;
  • 윤영심 (충북대학교 수의과대학 및 동물의학연구소) ;
  • 강신영 (충북대학교 수의과대학 및 동물의학연구소)
  • Bae, Su-Jung (College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University) ;
  • Kim, Jin-Won (College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University) ;
  • Yoon, Young-Sim (College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University) ;
  • Kang, Shien-Young (College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University)
  • 발행 : 2009.03.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Porcine reproductive and respiratory syndrome (PRRS) virus is the etiological agent of diseases characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRS virus is a small enveloped virus containing a positive-sense, single-stranded RNA genome. In the present study, ORF6 gene of Korean PRRS virus isolate, CNV, was cloned and expressed in baculovirus expression system. The ORF6 gene and expressed protein in the recombinant virus were confirmed by PCR/indirect fluorescence antibody (IFA) test and Western blotting, respectively. The recombinant protein with a molecular weight of approximately 24KDa was confirmed by Western blotting using His6 and PRRS virus-specific antiserum. Expressed ORF6 protein was applied for IFA to detect antibody against PRRS virus using field porcine sera. However, the sensitivity and specificity of developed IFA using expressed ORF6 protein were considerably low compared to those of commercial ELISA kit. This results suggest that IFA using expressed ORF6 protein could not be used as a diagnostic test for PRRS virus infection without further improvements.

키워드

참고문헌

  1. 박최규, 류영수, 이창희 등. 1998. 돼지 생식기호흡기증후군 바이러스항체 검색에 있어간접형광항체법(IFA)과 효소면역법(ELISA)의 진단효율 비교. 대한수의학회지 38: 314-318
  2. Albina E, Leforban Y, Baron T, et al. 1992. An enzymc linked immunosorbent assay (ELlSA) for the detection of antibody to porcine reproductive and respiratory syndrome virus. Ann Rec Vet 23: 167-176
  3. Bautista EM, Meulenberg JJ, Choi CS, et al. 1998. Structural polypeptidεs of the American (VR-2332) strain of porcine reproductivε andrεspiratory syndrome virus. Arch Virol 141: 357-1365 https://doi.org/10.1007/BF01718405
  4. Cavanagh D. 1997. Nidovirales: A new order comprising Coronaviridae and Arteriviridae. Arch Virol 142: 629-633
  5. Dea S, Gagnon CA, Mardassi H, et al. 2000. Current know ledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus. Arch Virol 145: 659-688 https://doi.org/10.1007/s007050050662
  6. Houben S, Callebaut P, Pensaert MB. 1995. Comparative study of a blocking enzyme linked immunosorbent assay and the immunoperoxidase monolayer assay for the detection of antibody to porcine reproductive and respiratory syndrome virus in pigs. J Virol Methods 51: 125-128 https://doi.org/10.1016/0166-0934(94)00135-4
  7. Kamata K, Shinozaki K, Okada M, et al. 2005. Expression and antigenicity of virus-like particles of norovirus and their application for detection of noroviruses in stool samples. J Med Virol 6: 129-136
  8. Keffaber KK. 1989. Reproductive failure of unknown etiology. Am Assoc Swine Pract Newsletter 1: 1-10
  9. Kim HS, Kwang J, Yoon IJ, et al. 1993. Enhanced replication of porcine reproductivε and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line. Arch Virol 133: 477-483 https://doi.org/10.1007/BF01313785
  10. Kreutz LC, Mengeling WL. 1997. Baculovirus expression and immunological detection of the major structural proteins of porcine reproductive and respiratory syndrome virus. Vet Microbiol 59: 1-13 https://doi.org/10.1016/S0378-1135(97)00142-9
  11. Perrin A, Albina E, Breard E, et al. 2007. Recombinant capri poxviruses expressing protein of bluetongue virus: evaluation of immune responses and protection in small ruminants. Vaccine 25: 6774-6783 https://doi.org/10.1016/j.vaccine.2007.06.052
  12. Qian P, Li X, Tong G, et al. 2003. High-level expεrssion of the ORF6 gene of porcine reproductive and respiratory syndrome virus (PRRSV) in Pichia pastoris. Virus Genes 27: 189-196 https://doi.org/10.1023/A:1025784712265
  13. Snijder EJ, Meulenberg JJ. 1998. The molecular biology of arterivirusεs. J Gen Virol 79: 961-979
  14. Sorensen KJ, Strandbygaard B, Botner A, et al. 1998. Blocking ELISA's for the distinction between antibodies against european and american strains of porcine reproductive and respiratory syndrome virus. Vet Microbiol 60: 169-177 https://doi.org/10.1016/S0378-1135(98)00159-X
  15. Stεwart ME, Bonne N, Shearer P, et al. 2007. Baculovirus expre, ssion of beak and feather disease virus (BFDV) capsid protein capable of self-assembly and haemagglutination. J Virol Methods 141: 181-187 https://doi.org/10.1016/j.jviromet.2006.12.011
  16. Towbin H, Staehelin T, Gordon J. 1985. Electrophoretic transfεr of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350-4354 https://doi.org/10.1073/pnas.76.9.4350
  17. Wensvoort G, Terpstra C, Laak EA, et al. 1991. Mystery swine disease in the Netherlands: The isolation of Lelystad virus. Vet Quart 13: 121-130 https://doi.org/10.1080/01652176.1991.9694296
  18. Yoon IJ, Joo HS, Christianson WT, et al. 1992. An indirect fluorεscent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera, J Vet Diagn Invest 4: 144-147 https://doi.org/10.1177/104063879200400205
  19. Yoon IJ, Joo HS, Goyal SM, et al. 1994. A modified serum neutralization test for the detection of antibody to porcine reproductive and respiratory syndrome virus in swine sera, J Vet Diahn Invest 6: 289-292
  20. Zimmerman JJ, Benfield DA, Murtaugh MP, et al. 2006. Porcine reproductive and respiratory syndrome virus (Por cine arterivirus). In: Disease of swine. Straw BE, Zimmerman JJ, D'Allaire S, Taylor DL. eds. 9th eds, Blackwell publishing, Oxford: 387-417