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Expression of the structural proteins of porcine reproductive and respiratory syndrome virus in Bm5 cells

  • 언어ENG
  • URLhttps://db.koreascholar.com/Article/Detail/290773
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

The porcine reproductive and respiratory syndrome virus (PRRSV) has six structural proteins which encoded by ORFs 2 to 7 are designated as GP2, 3, 4, 5, M and N, repectively. In this study, we determined the expression of each protein using novel transfer vector, pBmKSK4 which has the polyhedrin promoter of BmNPV and 6xHis tag. The recombinant transfer vector was co-transfected into Bm5 cells along with bBpGOZA DNA. Recombinant virus was purified by plaque assay and amplified in Bm5 cells. Expression of each protein was identified by SDS-PAGE and Western blot analysis using anti-6xHis monoclonal antibody. The expression levels of the structural proteins in Bm5 cells were stronger than the expression system using pBacPAK9 transfer vector in Sf21 cells. As expected, GP5 was expressed at low levels from its structural properties and its toxicity for cells. In addition, each recombinant protein was purified using Ni-NTA spin columns. The ability to produce each protein in the baculovirus system indicates that these could be major candidates for the development of a vaccine against PRRSV.

저자
  • Soo Dong Woo(Department of Plant Medicine, Chungbuk National University)
  • In Joon Yoon(Choong-Ang Vaccine Laboratory)
  • Young In Kim(AgroLife Research Institute (ARI), Dongbu HiTek Co. Ltd.)
  • Jae Su Kim(AgroLife Research Institute (ARI), Dongbu HiTek Co. Ltd.)
  • Sung Sik Yoo(Choong-Ang Vaccine Laboratory)
  • Byung Rae Jin(College of Natural Resources and Life Science, Dong-A University)
  • Yeon Ho Je(School of Agricultural Biotechnology, Seoul National University)
  • Jong Yul Roh(School of Agricultural Biotechnology, Seoul National University)
  • Kwang Sik Lee(College of Natural Resources and Life Science, Dong-A University)
  • Jae Young Choi(School of Agricultural Biotechnology, Seoul National University)
  • Hyun Na Koo(Department of Plant Medicine, Chungbuk National University)
  • Jae Kyung Lee(Department of Plant Medicine, Chungbuk National University)
  • Jeong Mi Oh(Department of Plant Medicine, Chungbuk National University)