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| PTD-hEGF融合蛋白在大肠杆菌中的表达及活性分析 | |
| 引用本文： | 智庆文,王淑豪,张凤英,李仕贵,孙曼霁.PTD-hEGF融合蛋白在大肠杆菌中的表达及活性分析[J].生物工程学报,2007,23(4):589-592. |
| 作者姓名： | 智庆文 王淑豪 张凤英 李仕贵 孙曼霁 |
| 作者单位： | 1. 军事医学科学院毒物药物研究所,北京,100850;四川农业大学教育部作物遗传资源和发展重点实验室,雅安,625014;中国人民解放军防化指挥工程学院,北京,102205 2. 军事医学科学院毒物药物研究所,北京,100850 3. 中国人民解放军防化指挥工程学院,北京,102205 4. 四川农业大学教育部作物遗传资源和发展重点实验室,雅安,625014 |
| 基金项目： | 致谢 MALDI-T0F-MS分析结果是在军事医学科学院仪器测试分析中心王杰老师的帮助和指导下完成的,在此表示感谢. |
| 摘 要： | 为获得hEGF在大肠杆菌中的高效表达,构建了pPTD-hEGF原核表达载体。将其转化大肠杆菌BL21(DE3),经0.3mmol/LIPTG诱导,PTD-hEGF融合蛋白进行了有效表达,表达产物主要形成了包涵体。经Ni2 -NTA亲和层析纯化,融合蛋白的纯度达99.5%。MALDI-TOF-MS分析证明表达的融合蛋白氨基酸序列完全正确。PTD-hEGF融合蛋白能明显促进HEK-293细胞的分裂和生长。 |
| 关 键 词： | PTD-hEGF融合蛋白 原核表达 HEK-293细胞 |
| 文章编号： | 1000-3061（2007）04-0589-04 |
| 修稿时间： | 2006-11-16 |
PTD-hEGF Fusion Protein-Gene Expression and Function Analysis |
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| ZHI Qing-Wen,WANG Shu-Hao,ZHANG Feng-Ying,LI Shi-Gui and SUN Man-Ji.PTD-hEGF Fusion Protein-Gene Expression and Function Analysis[J].Chinese Journal of Biotechnology,2007,23(4):589-592. | |
| Authors: | ZHI Qing-Wen WANG Shu-Hao ZHANG Feng-Ying LI Shi-Gui and SUN Man-Ji |
| Institution: | 1 Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Belting 100850, China 2 Key Laboratory of Crop Genetic Resource and Improvement of Ministry of Education, Sichuan Agrlcaltural University, Yaan 625014, China 3 Command and Engineering College of Chemical Defense of Chinese PLA ,Belling 102205, China |
| Abstract: | To produce membrane-permeable human epidermal growth factor (hEGF),a pPTD-hEGF prokaryocyte expression vector was constructed and transformed into E. coli BL 21 (DE3). The PTD-hEGF fusion protein was induced by 0.3mmol/L of IPTG expressed in the form of inclusion body with an yield of 40% of the total protein in the cells,and then purified by Ni~ 2 -NTA affinity chromatography. The SDS-PAGE analysis showed a single fusion protein band with a molecular weight of 16kD. The amino acid sequence was checked by MALDI-TOF-MS analysis. The genetic engineering PTD-hEGF fusion protein obviously promoted the proliferation and growth of the HEK-293 cells in vitro. |
| Keywords: | PTD-hEGF fusion protein prokaryocyte expression HEK-293 cell |
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