Korean Journal of Veterinary Research 2004;44(2):251-257.
Production of expressed protein from cloned ShigatoxinG 2e gene and Receptor Binding Affinity of the toxin
Bun-youn Dong1, Sang-Hyun Kim2, Yeong-Il Kim3, Hyun-Ho Cho3, Woo-won Lee4, Kon-Sup Kim5, Ho-Jo Kang5, Yong-Hwan Kim5
1Korea Green Cross Co.
2Department of Pathobiology, Ontario Veterinary College, University of Guelph
3National Veterinary Research and Quarantine Service
4Pusan Institute of Health and Environment
5College of Veterinary Medicine, Research Institute of Life Science,Kyeongsang National University
재조합 Shigatoxin 2e 유전자의 발현단백 생산 및 독소의 수용체 결합 친화성 확인
동분연1, 김상현2, 김영일3, 조현호3, 이우원4, 김곤섭5, 강호조5, 김용환5
2캐나다 구엘프대학교 수의과대학
3수의과학검역원 부산지소
4부산시 보건환경 연구원
5경상대학교 수의과대학 생명과학연구소
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.
Key Words: Shigatoxin 2e gene, expression, cytotoxicity, globotetraosyl ceramide, receptor binding ELISA
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