| 程 虹,金庆文,姚 娟,邓晓萱,万 琪,丁新生,季晓辉.血管内皮生长因子在大肠杆菌中表达?提纯及功能测定[J].南京医科大学学报,2008,28(1):33~36 |
| 血管内皮生长因子在大肠杆菌中表达?提纯及功能测定 |
| Expression,purification,and characterization of VEGF in E. Coli |
| 投稿时间：2007-09-09 |
| DOI：10.7655 |
| 中文关键词: 血管内皮生长因子 PET-32a(+)表达载体 Neuropilin-1 |
| 英文关键词: VEGF PET32a expression vector Neuropilin-1 |
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| 中文摘要: |
| 目的:利用细菌表达系统合成大量重组血管内皮生长因子(VEGF)蛋白?方法:将VEGF165 cDNA克隆于表达载体PET-32a(+)后转染BL-21株表达重组VEGF蛋白,用分离His蛋白的层析柱提纯重组VEGF蛋白?结果:重组VEGF蛋白经氨基酸序列测定证实?Western blot测定发现His蛋白?功能测定发现重组VEGF蛋白可阻断细胞表面Neuropilin-1的表达?结论:上述表达系统制备大量具有生物活性的重组VEGF蛋白 |
| 英文摘要: |
| Objective:To make recombinant vascular endothelial growth factor(VEGF) in E.Coli. Methods:The VEGF cDNA was cloned and put into the bacterial expression system PET32a(Novagen) in frame with histidine tag. After introducing into the BL21 (DE3) E coli host,recombinant VEGF165 protein was produced and purified by denaturing condition on the His-bind resin. Results:The recombinant protein was confirmed by AA sequencing. The reconstituted VEGF165 was recognized by its receptor Neuropilin-1 and could obstruct the expression of Neuropilin-1 in cell curface. Conclusion:pET32a/human VEGF165 expression vector was transfected into E.coli BL21(DE3) and produced a lot of functional recombinant human VEGF165. |
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