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| TAT-EGFP融合蛋白表达载体的构建及表达 | |
| 引用本文： | 王海珍,许予明,赵莘瑜,陈奎生,李惠翔,张云汉.TAT-EGFP融合蛋白表达载体的构建及表达[J].郑州大学学报(医学版),2007,42(3):461-463. |
| 作者姓名： | 王海珍 许予明 赵莘瑜 陈奎生 李惠翔 张云汉 |
| 作者单位： | 1. 河南省肿瘤病理重点实验室,郑州大学第一附属医院病理科,郑州,450052;郑州大学第一附属医院神经内科,郑州,450052 2. 郑州大学第一附属医院神经内科,郑州,450052 3. 河南省肿瘤病理重点实验室,郑州大学第一附属医院病理科,郑州,450052 |
| 基金项目： | 河南省杰出青年科学基金 |
| 摘 要： | 目的:构建TAT-EGFP原核表达载体,在E.coli BL21中高效表达并纯化.方法:人工合成编码TAT蛋白转导区的DNA片段,插入载体pET28a后得到pET28a-TAT重组质粒,再连接绿色荧光蛋白(EGFP)基因,组成pET28a-TAT-EGFP重组表达子.转化大肠杆菌,IPTG诱导TAT-EGFP融合蛋白表达.表达产物用十二烷基硫钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定,组氨酸亲和层析柱纯化融合蛋白.结果及结论:成功地构建了TAT-EGFP融合蛋白的原核表达载体,在诱导后获得了高效表达并纯化,为进一步研究TAT的蛋白转导作用奠定了基础. |
| 关 键 词： | TAT 蛋白转导结构域 融合蛋白 |
| 收稿时间： | 2006-09-29 |
| 修稿时间： | 2006-09-29 |
Construction and expression of TAT-EGFP fusion protein expression vector |
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| WANG Haizhen,XU Yuming,ZHAO Xinyu,CHEN Kuisheng,LI Huixiang,ZHANG Yunhan.Construction and expression of TAT-EGFP fusion protein expression vector[J].Journal of Zhengzhou University: Med Sci,2007,42(3):461-463. | |
| Authors: | WANG Haizhen XU Yuming ZHAO Xinyu CHEN Kuisheng LI Huixiang ZHANG Yunhan |
| Institution: | 1 Henan Key Laboratory of Tumor Pathology ; Department of Pathology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 ;2 Department of Neurology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 |
| Abstract: | Aim: To construct an expression vector of TAT-EGFP and express the fusion protein in E.coli BL21.Methods:A synthesized DNA fragment encoding TAT protein transduction domain was inserted into pET28a vector, then EGFP gene was ligated to construct the expression vector of pET28a-TAT-EGFP. The recombinant vector was transformed into E.coli BL21 and induced with IPTG. The highly expressed TAT-EGFP was purified by affinity chromatography.Results and Conclusion: pET28a-TAT-EGFP is successfully constructed and fusion protein TAT-EGFP is successfully expressed in prokaryotic cells, which lays the foundation for studying the protein transduction of TAT. |
| Keywords: | TAT protein transduction domain fusion protein |
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