| CTB-InsB融合基因在大肠杆菌中的表达研究 |
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| 引用本文： | 徐元力,童富淡,张耀洲.CTB-InsB融合基因在大肠杆菌中的表达研究[J].浙江理工大学学报,2007,24(4):471-474. |
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| 作者姓名： | 徐元力 童富淡 张耀洲 |
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| 作者单位： | 浙江理工大学生命科学学院生物化学研究所,杭州,310018 |
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| 基金项目： | 国家高技术研究发展计划(863计划);国家重点基础研究发展计划(973计划);浙江省自然科学基金 |
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| 摘 要： | 构建了一种霍乱毒素B亚基（CTB）与人胰岛素B链（InsB）的融合基因，并克隆入表达载体pGEX-4T-1中，命名为pGEX—CTB-InsB。该融合蛋白在大肠杆菌中的表达产物以包涵体形式存在，分子量约为43kDa。包涵体经溶解和复性后，使用GSTrapFF亲和层析柱纯化得到融合蛋白。经SDS-PAGE和Western免疫印迹分析证实纯化蛋白为重组目的蛋白。
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| 关 键 词： | CTB-InsB融合基因 诱导表达 纯化 |
| 文章编号： | 1673-3851（2007）04-0471-04 |
| 收稿时间： | 2007-03-07 |
| 修稿时间： | 2007-03-07 |
Genetic Fusion of Cholera Toxin B Subunit and Human Insulin B Chain and Its Expression in E.Coli |
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| XU Yuan-li,TONG Fu-dan,ZHANG Yao-zhou.Genetic Fusion of Cholera Toxin B Subunit and Human Insulin B Chain and Its Expression in E.Coli[J].Journal of Zhejiang Sci-tech University,2007,24(4):471-474. |
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| Authors: | XU Yuan-li TONG Fu-dan ZHANG Yao-zhou |
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| Affiliation: | School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China |
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| Abstract: | In the study,the construction of genetic fusion of the cholera toxin B subunit and human insulin B chain is described(In this fusion protein,InsB was fused to the C-terminus of a CTB monmer).The resulting fusion gene is cloned into pGEX-4T-1 expression vector and is named aspGEX-CTB-InsB.The resulting fusion protein is expressed in Escherichia coli and isolated as cytoplasmic inclusion bodies that are then dissolved and assembled in vitro.After a serial process and the GSTrap FF Affinity chromatography,the authors get the purified protein fusing with GST at the N-terminus of CTB,which is about 43 kDa.SDS-PAGE and Western blot analyses show that the purified protein is the aim protein. |
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| Keywords: | CTB-InsB genetic fusion orotein Inducible exoression Purification |
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