| 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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| 引用本文： | 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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| 作者姓名： | 赵 剑 徐俊杰 缪 静 李 冰 杨秀旭 宋小红 陈 薇 |
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| 作者单位： | 1. 军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京,100071
2. 烟台师范大学生命科学院,烟台,264025 |
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| 基金项目： | 国家自然科学基金资助项目(30300016). |
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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 .
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| 关 键 词： | 炭疽杆菌，炭疽毒素受体，蛋白质表达，毕赤酵母 |
Expression, Purification and Characterization of The Recombinant Excellular Portion of Anthrax Toxin Receptor |
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| ZHAO Jian,XU Jun-Jie,MIAO Jing,LI Bing,YANG Xiu-Xu,SONG Xiao-Hong and CHEN Wei.Expression, Purification and Characterization of The Recombinant Excellular Portion of Anthrax Toxin Receptor[J].Progress In Biochemistry and Biophysics,2005,32(7):657-661. |
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| Authors: | ZHAO Jian XU Jun-Jie MIAO Jing LI Bing YANG Xiu-Xu SONG Xiao-Hong and CHEN Wei |
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| Institution: | State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;School of Life Science, Yantai Normal University, Yantai 264025, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China |
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| 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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| Keywords: | Bacillus anthracis anthrax toxin receptor protein expression Pichia pastoris |
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