| 大肠杆菌硫氧还蛋白thioredoxin基因的克隆及表达 |
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| 引用本文： | 席慧,刘凤华,李永海,徐燕,高音. 大肠杆菌硫氧还蛋白thioredoxin基因的克隆及表达[J]. 首都师范大学学报(自然科学版), 2004, 25(4): 62-65,70 |
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| 作者姓名： | 席慧 刘凤华 李永海 徐燕 高音 |
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| 作者单位： | 首都师范大学生物系,北京,100037;首都师范大学生物系,北京,100037;首都师范大学生物系,北京,100037;首都师范大学生物系,北京,100037;首都师范大学生物系,北京,100037 |
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| 基金项目： | 国家自然科学基金资助项目 (No .3 980 0 0 2 8) |
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| 摘 要： | 以大肠杆菌XL1blue为模板，通过PCR技术扩增大肠杆菌硫氧还蛋白基因，并将目的基因分别连接到克隆载体pUC18和表达载体pTrcHisC上，构建重组质粒．重组质粒pTrcHisC-TRX在大肠杆菌中高效表达，最后利用固定化金属螫合亲和层析技术(IMAC)获得较纯的目的蛋白，为进一步研究硫氧还蛋白的功能及其应用提供了条件．
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| 关 键 词： | 蛋白质二硫键异构酶 大肠杆菌硫氧还蛋白 固定化金属螯合亲和层析 诱导表达 |
Cloning and Expression of Escherichia coli Thioredoxin |
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| Xi Hui Liu Fenghua Li Yonghai Xu Yan Gao Yin. Cloning and Expression of Escherichia coli Thioredoxin[J]. Journal of Capital Normal University(Natural Science Edition), 2004, 25(4): 62-65,70 |
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| Authors: | Xi Hui Liu Fenghua Li Yonghai Xu Yan Gao Yin |
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| Abstract: | By using the Polymerase Chian Reaction (PCR) technique,the 330?bp DNA fragment of trx gene was amplified from the Escherichia coli XL1blue cell.The gene was inserted into expression vector pTrcHisC after cloned into plasmid pUC18 for sequencing.Protein expression was induced by IPTG,and the recombinant TRX was purified by using TALON metal affinity resin column chromatography.Our experiments make a base for the further study of this protein's configuration and function. |
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| Keywords: | protein disulfide isomerase Escherichia coli thioredoxin immobilized metal affinity chromatography (IMAC) inducing expression. |
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