| 人血管内皮细胞生长因子在大肠杆菌中的表达 |
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| 引用本文： | 王雅梅,祁雅慧,司杨,殷红,武文琦. 人血管内皮细胞生长因子在大肠杆菌中的表达[J]. 首都医科大学学报, 2003, 24(2): 111-114 |
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| 作者姓名： | 王雅梅 祁雅慧 司杨 殷红 武文琦 |
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| 作者单位： | 首都医科大学实验中心(王雅梅，祁雅慧，司杨，殷红)，首都医科大学实验中心(武文琦) |
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| 基金项目： | 北京市科委科技合同资金(954024800) |
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| 摘 要： | 应用DNA重组技术,将编码人血管内皮细胞生长因子(hVEGF)的基因插入到原核表达载体pET32M中,与载体中的硫氧还蛋白(相对分子质量约1.4万)基因融合。经双酶切鉴定、序列测定及工程菌初步表达分析,表明该重组质粒在大肠杆菌BL21中有明显表达。表达的硫氧还蛋白-VEGF融合蛋白的相对分子质量约为3.2万,主要以包涵体形式存在,也有少量重组蛋白为可溶性蛋白,Western印迹实验和生物活性分析证实可溶性的重组蛋白具有hVEGF的抗原性和促血管生长活性。
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| 关 键 词： | 血管内皮细胞生长因子 融合蛋白 诱导表达 |
| 收稿时间： | 2002-09-20 |
| 修稿时间： | 2002-09-20 |
Expression of Vascular Endothelial Growth Factor in E.coli |
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| Wang Yamei,Qi Yahui,Si Yang,Yin Hong,Wu Wenqi. Expression of Vascular Endothelial Growth Factor in E.coli[J]. Journal of Capital Medical University, 2003, 24(2): 111-114 |
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| Authors: | Wang Yamei Qi Yahui Si Yang Yin Hong Wu Wenqi |
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| Affiliation: | Wang Yamei,Qi Yahui,Si Yang,Yin Hong,Wu Wenqi Department of Experiment Center,Capital University of Medical Sciences |
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| Abstract: | The DNA sequence coding for human Vascular Endothelial Growth Factor (hVEGF) was inserted into the down stram of thioredoxin gene in pET32M to generate thioredoxin-VEGF fusion protein expression vector pET32M/V. After identification and transformation, the thioredoxin-VEGF fusion protein with the molecular weight of 32 000 was detected by SDS-PAGE analysis and Western blotting after induced expression in E. coli BL21. The recombinant proteins were recovered as inclusion bodies, but there were also some soluble recombinant proteins. Chick Chorioallantoic Membrane (CAM) bioassay showed that recombinant protein had biological activities of hVEGF. |
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| Keywords: | vascular endothelial growth factor fusion protein induced expression |
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