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| 白喉毒素突变体CRM197原核表达载体构建及其蛋白表达 | |
| 引用本文： | 肖钾钙镁,王素英,魏文进,张静飞,林福玉.白喉毒素突变体CRM197原核表达载体构建及其蛋白表达[J].天津师范大学学报(自然科学版),2012,32(3):78-80. |
| 作者姓名： | 肖钾钙镁 王素英 魏文进 张静飞 林福玉 |
| 作者单位： | 1. 北京民海生物科技有限公司研发中心结合疫苗北京市重点实验室,北京102600;天津商业大学生物技术与食品科学学院,天津300134 2. 天津商业大学生物技术与食品科学学院,天津,300134 3. 北京民海生物科技有限公司研发中心结合疫苗北京市重点实验室,北京,102600 |
| 摘 要： | 将合成的CRM197基因片段克隆到表达载体pBAD-DEST49中，转化到大肠杆菌菌株（ATCCMC1061）．经阿拉伯糖诱导，CRM197获得高效表达．目的蛋白主要以包涵体形式存在，变性、复性后经DEAE阴离子交换后获得高于95％纯度的CRM197样品．用该样品做Westernblotting鉴定后，能得到单一的特异性条带．本实验的表达策略，为CRM197进一步生产及应用奠定基础． |
| 关 键 词： | CRM197 白喉毒素 大肠杆菌 |
Expression of diphtheria toxin mutant CRM197 by construction of prokaryotic vector in Escherichia coli |
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| XIAO Jia-gai-mei , WANG Su-ying , WEI Wen-jin , ZHANG Jing-fei , LIN Fu-yu.Expression of diphtheria toxin mutant CRM197 by construction of prokaryotic vector in Escherichia coli[J].Journal of Tianjin Normal University(Natural Science Edition),2012,32(3):78-80. | |
| Authors: | XIAO Jia-gai-mei WANG Su-ying WEI Wen-jin ZHANG Jing-fei LIN Fu-yu |
| Institution: | 1.Beijing Key Laboratory of Conjugate Vaccine,Beijing Minhai Biological Science and Technology Co.Ltd,Beijing 102600,China;2.College of Biotechnology and Food Science,Tianjin University of Commerce,Tianjin 300134,China |
| Abstract: | The gene of CRM197(cross reacting material 197,CRM197) is synthesized and then expressed in Escherichia coli(ATCCMC1061) driven by pBAD-DEST49.CRM197 is efficiently expressed by Arabinose induced.The recombinant protein is expressed mainly as inclusion body.After denaturation,renaturation and purification by DEAE anion-exchange column,CRM197 with the purity higher than 95% is obtained.For Western blotting,the protein can get single specific bands.Given these characteristics,the CRM197 expressed in Escherichia coli is suitable for further research. |
| Keywords: | CRM197 diphtheria toxin Escherichia coli |
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