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

作　　者：程虹[1] 金庆文[1] 姚娟[1] 邓晓萱[1] 万琪[1] 丁新生[1] 季晓辉[2] 

机构地区：[1]南京医科大学第一附属医院神经内科,江苏南京210029 [2]南京医科大学微生物教研室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2008年第1期33-36,共4页Journal of Nanjing Medical University(Natural Sciences)

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VEGF165cDNA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。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 VEGFI6s expression vector was transfected into E.coli BL21 （DE3） and produced a lot of functional recombinant human VEGF165.

关 键 词：血管内皮生长因子 PET-32a(+)表达载体 NEUROPILIN-1 

分 类 号：R392.11[医药卫生—免疫学]