Analysis of diphtheria toxin receptor by molecular cloning.
Analysis of diphtheria toxin receptor by molecular cloning.
批准号:
62570189
负责人:
MEKADA Eisuke
金额:
$1.41万
依托单位国家:
日本
项目类别:
Grant-in-Aid for General Scientific Research (C)
财政年份:
1987
资助国家:
日本
项目状态:
已结题
起止时间:
1987 至 1988
中文摘要
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英文摘要
The aim of this project is to understand the entry mechanism of diphtheria toxin into cytosol. The strategy of this project is as follows: (1)Identification of diphtheria toxin receptor in membrane fractions of cultured cells. (2)Purification of the receptor. (3)Isolation of monoclonal antibodies against diphtheria toxin receptor. (4)cDNA cloning of the receptor using monoclonal antobodies.In 1987, we identified diphtheria toxin receptor as 14.5Kd protein in lysate of Vero cell membrane using CRM197 protein, a mutant protein of diphtheria toxin. Then the receptor was purified by a 10,000-folds by sequential chromatography. In this year, we tried to isolate hybridomas producing antibodies directed to diph-theria toxin receptor. Several clones were isolated. An antibody produced from one of these clones inhibited the binding of diphtheria toxin to the receptor. This antibody specefically precipitates the 14.5Kd protein in the fraction of partially purified receptor. Now we tried to isolate cDNA of the receptor using these monoclonal antibodies.
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Eisuke Mekada: "Hybrid Molecules in Handbook of Natural Toxin,vol.IV.," Mercel Dekker Inc., 36 (1988)
Eisuke Mekada:“天然毒素手册中的混合分子,第 IV 卷”,Mercel Dekker Inc.,36 (1988)
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目加田英輔: 微生物. 3. 66-72 (1987)
Eisuke Mekada:微生物。3. 66-72 (1987)
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Kenji Kohno: Exptl.Cell Res.172. 54-64 (1987)
Kenji Kohno:Exptl.Cell Res.172。
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Eisuke Mekada: "Identification of diphtheria toxin receptor and a nonproteinous diphtheria toxin-binding molecule in Vero cell membrane." J. Cell Biol.107. 511-519 (1988)
Eisuke Mekada:“鉴定 Vero 细胞膜中的白喉毒素受体和非蛋白白喉毒素结合分子。”
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