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Influence of insect cells and media conditions for the expression of glycoprotein E2 of classical swine fever virus

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  • URLhttps://db.koreascholar.com/Article/Detail/290425
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한국응용곤충학회 (Korean Society Of Applied Entomology)
초록

The Classical Swine Fever Virus (CSFV) is a member of the Pestivirus genus of the Flaviviridae. The genome of CSFV is a positive single-stranded RNA molecule 12.3 kb and contains a single large open reading frame (ORF). The polyprotein composed of eight nonstructural and four structural proteins (nucleocapsid protein C and three envelope glycoprotein E0, E1 and E2). E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. To determine the characteristics of the CSFV, LOM strain, we investigated the nucleotide sequence of the glycoprotein E0, E1 and E2. Comparison of the LOM with the other strains revealed nucleotide sequence identity ranging from 97 to 98%. Expression of the glycoprotein E2 was identified by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies in Sf21 cells. The expression levels of glycoprotein E2 were observed from day 3 and 5 days maximum. In addition, its expression efficiency by media and cell line was investigated. The result showed that High-Five cells and Grace’s insect media for Sf21 were the best conditions for the expression of the glycoprotein E2.

저자
  • Sung Min Bae(Department of agricultural biology, Chungbuk National University)
  • Hyun Na Koo(Department of agricultural biology, Chungbuk National University)
  • Tae Young Shin(Department of agricultural biology, Chungbuk National University)
  • Jae Bang Choi(Department of agricultural biology, Chungbuk National University)
  • Jae Young Choi(School of Agricultural Biotechnology, Seoul National University, Research Institute for Agriculture and Life Sciences, Seoul National University)
  • Kwang Sik Lee(College of Natural Resources and Life Science, Dong-A University)
  • Jong Yul Roh(School of Agricultural Biotechnology, Seoul National University)
  • Yeon Ho Je(School of Agricultural Biotechnology, Seoul National University)
  • Byung Rae Jin(College of Natural Resources and Life Science, Dong-A University)
  • Sung Sik Yoo(Choong-Ang Vaccine Laboratory)
  • Jae Su Kim(AgroLife Research Institute (ARI), Dongbu HiTek Co. Ltd.)
  • Young In Kim(AgroLife Research Institute (ARI), Dongbu HiTek Co. Ltd.)
  • In Joong Yoon(Choong-Ang Vaccine Laboratory)
  • Soo Dong Woo(Department of agricultural biology, Chungbuk National University)