| F3重组毒素的克隆表达及纯化 |
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| 引用本文： | 朱海兵,黄轶,曾昭淳.F3重组毒素的克隆表达及纯化[J].科技导报(北京),2009,27(12). |
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| 作者姓名： | 朱海兵 黄轶 曾昭淳 |
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| 作者单位： | 重庆医科大学生物化学及分子生物学教研室,重庆,400016
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| 摘 要： | 以pEKH-F3质粒为模板,PCR扩增F3片段,将F3插入克隆质粒PQE80L,转化大肠杆菌,筛选,获得含有F3的PQE80L重组载体的克隆.提取绿脓杆菌菌株染色体DNA为模板,PCR扩增绿脓杆菌外毒素A催化区,PE40.然后将PQE80-F3和PE40重组,得到表达PQE80L-F3-PE40载体.转化至大肠杆菌DH5a,BL21,表达融合蛋白F3-PE40.结果显示,大肠杆菌表达了融合蛋白F3-PE40,表达的融合蛋白量大概占菌体总体蛋白量的20%.通过质粒提取、PCR扩增构建F3-PE40表达载体转化大肠杆菌,成功地表达了融合蛋白F3-PE40,为大规模表达、纯化F3-PE40的进一步功能奠定了基础.
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| 关 键 词： | 融合基因 克隆 表达 绿脓杆菌外毒素A催化区 |
The Cloning, Expression and Purification of F3 Recombinant Toxin |
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| ZHU Haibing,HUANG Yi,ZENG Zhaochun.The Cloning, Expression and Purification of F3 Recombinant Toxin[J].Science & Technology Review,2009,27(12). |
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| Authors: | ZHU Haibing HUANG Yi ZENG Zhaochun |
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| Institution: | ZHU Haibing,HUANG Yi,ZENG Zhaochun Department of Biochemistry , Molecular Biology,Chongqing Medical University,Chongqing 400016,China |
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| Abstract: | F3 is inserted into PQE80L carrier, after being amplified by PCR, with the plasmid pEKH -F3 as the template, to obtain PQE80L -F3 recombinated plasmid. With extracted pseudomonas aeruginosa strain chromosome DNA as the template, PE40, the pseudomonas aeruginosa exotoxin A catalysis area, is amplified by PCR. Then PQE80 -F3 is recombinated with PE40, to obtain PQE80L-F3-PE40 carrier. The DH5a and BL21 are obtained by transformation. The results show that F3 -PE40 fusion protein is expressed and the quantity ... |
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| Keywords: | fusion gene clone expression pseudomonas aeruginosa exotoxin A catalysis area |
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