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

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)
Publication Information
Korean Journal of Veterinary Service / v.32, no.1, 2009 , pp. 19-25 More about this Journal
Abstract
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.
Keywords
PRRS virus; ORF6; Baculovirus expression;
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1 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   DOI   ScienceOn
2 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   DOI   ScienceOn
3 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   DOI   ScienceOn
4 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   DOI   ScienceOn
5 박최규, 류영수, 이창희 등. 1998. 돼지 생식기호흡기증후군 바이러스항체 검색에 있어간접형광항체법(IFA)과 효소면역법(ELISA)의 진단효율 비교. 대한수의학회지 38: 314-318
6 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   DOI   ScienceOn
7 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   DOI   PUBMED
8 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
9 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
10 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
11 Snijder EJ, Meulenberg JJ. 1998. The molecular biology of arterivirusεs. J Gen Virol 79: 961-979   PUBMED
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   DOI   ScienceOn
13 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
14 Keffaber KK. 1989. Reproductive failure of unknown etiology. Am Assoc Swine Pract Newsletter 1: 1-10
15 Cavanagh D. 1997. Nidovirales: A new order comprising Coronaviridae and Arteriviridae. Arch Virol 142: 629-633   PUBMED
16 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   DOI   ScienceOn
17 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   DOI   ScienceOn
18 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   DOI   ScienceOn
19 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   DOI   ScienceOn
20 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   DOI   PUBMED   ScienceOn