• Title, Summary, Keyword: globotetraosyl ceramide

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Quantitative assay of glycosphingolipid and determination of binding affinity for Shigatoxin 2e in pig (돼지의 일령에 따른 glycosphingolipid의 정량 및 Shigatoxin 2e에 대한 친화성 확인)

  • Dong, Bun-youn;Kim, Sang-Hyun;Cho, Hyun-Ho;Kim, Yeong-Il;Lee, Woo-won;Kim, Kon-Sup;Kang, Ho-Jo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.233-239
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    • 2004
  • This study was designed to quantitative assay of cellular receptor globotriosyl ceramide($Gb_3$) and globotetraosyl ceramide($Gb_4$) and to determine binding affinity of the receptors for Shigatoxin 2e in difference with day olds of pig. The amount of $Gb_3$ and $Gb_4$ extracted from each swine tissue were detected in order of spleen, ileocecum, kidney, and jejunum. The amount of $Gb_3$ and $Gb_4$ extracted from each swine ileocecum and jejunum in different day old swine were detected in order of 24, 6, 98, 12 and 35, but the difference of amount was not significant. The binding affinity of $Gb_4$ extracted from ileocecum and jejunum with different day old swine for Stx2e were shown that the extracts of 24, 12, and 35 days old swine were higher significantly than that of 6 and 98 days old swine.

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;Kim, Sang-Hyun;Kim, Yeong-Il;Cho, Hyun-Ho;Lee, Woo-won;Kim, Kon-Sup;Kang, Ho-Jo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.251-257
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    • 2004
  • 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.