Production of expressed protein from cloned ShigatoxinG 2e gene and Receptor Binding Affinity of the toxin

재조합 Shigatoxin 2e 유전자의 발현단백 생산 및 독소의 수용체 결합 친화성 확인

  • Dong, Bun-youn (Korea Green Cross Co.) ;
  • Kim, Sang-Hyun (Department of Pathobiology, Ontario Veterinary College, University of Guelph) ;
  • Kim, Yeong-Il (National Veterinary Research and Quarantine Service) ;
  • Cho, Hyun-Ho (National Veterinary Research and Quarantine Service) ;
  • Lee, Woo-won (Pusan Institute of Health and Environment) ;
  • Kim, Kon-Sup (College of Veterinary Medicine, Research Institute of Life Science,Kyeongsang National University) ;
  • Kang, Ho-Jo (College of Veterinary Medicine, Research Institute of Life Science,Kyeongsang National University) ;
  • Kim, Yong-Hwan (College of Veterinary Medicine, Research Institute of Life Science,Kyeongsang National University)
  • 동분연 (녹십자백신연구소) ;
  • 김상현 (캐나다 구엘프대학교 수의과대학) ;
  • 김영일 (수의과학검역원 부산지소) ;
  • 조현호 (수의과학검역원 서울지소) ;
  • 이우원 (부산시 보건환경 연구원) ;
  • 김곤섭 (경상대학교 수의과대학 생명과학연구소) ;
  • 강호조 (경상대학교 수의과대학 생명과학연구소) ;
  • 김용환 (경상대학교 수의과대학 생명과학연구소)
  • Accepted : 2004.04.17
  • Published : 2004.06.30

Abstract

This study was designed to determine optimal condition for expression of cloned Shigatoxin2e(Stx2e) gene from transformed E. coli PED18, to compare the cytotoxicity titer between cloned Stx2e and Stx2e from original strain, and to confirm of receptor binding affinity of Stx2e for use of development of receptor binding ELISA to detect of Stx2e. The optimum composition of medium for expression of Stx2e gene in E.coli host-vector system was definded as the medium containing 0.5% glucose and 0.5 mM IPTG. The cytotoxicity titer of expressed Stx2e for Vero cell was 1000 fold higher than that of Stx2e from original strain AY93258. The binding affinity of Stx2e to receptor globotetraosyl ceramide($Gb_4$) was confirmed by immunobloting.

Keywords

References

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