炭疽毒素受体胞外区在毕赤酵母中分泌表达、纯化与活性鉴定
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国家自然科学基金资助项目(30300016).


Expression, Purification and Characterization of The Recombinant Excellular Portion of Anthrax Toxin Receptor
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This work was supported by a grant from The National Natural Sciences Fundation of China (303300016).

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    摘要:

    使用分泌型表达载体,实现了重组炭疽毒素受体胞外区 (rATR(CMG2)-EXCELL) 在毕赤酵母 KM71H 培养物上清中的分泌表达 . 表达量约占培养物上清总蛋白质的 20%. 经过螯合柱初步纯化,每升诱导培养物可获得约 1 mg 电泳纯的 rATR(CMG2)-EXCELL. 体外与配基 PA 结合试验和细胞保护试验显示, rATR(CMG2)-EXCELL 具有很好的生物活性 . rATR(CMG2)-EXCELL 的成功表达为今后研究炭疽毒素受体的作用机理、发展新型炭疽治疗药物打下基础 .

    Abstract:

    The gene fragment encoding ATR(CMG2)-EXCELL (excellular potion of anthrax toxin receptor/capillary morphogenesis factor) was cloned into a secretory expression plasmid and then expressed in media supernant of Pichia pastoris. The recombinant rATR(CMG2)-EXCELL expressed was about 20% of the total proteins in media supernant. About 1 mg electrophoresis purity rATR(CMG2)-EXCELL could be obtained after the purification of 1 L culure using chelating column. In vitro binding activity analysis and cell protection experiments have shown that rATR(CMG2)-EXCELL has an excellent biological activity. The successful expression of rATR(CMG2)-EXCELL has placed a solid foundation for the research on binding mechanism of ATR and PA(protective antigen) and developing new cure for anthrax.

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赵 剑,徐俊杰,缪 静,李 冰,杨秀旭,宋小红,陈 薇.炭疽毒素受体胞外区在毕赤酵母中分泌表达、纯化与活性鉴定[J].生物化学与生物物理进展,2005,32(7):657-661

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