重组血管内皮细胞生长因子在大肠杆菌的高效表达  

High level expression of recombinant VEGF in Escherichia Coli

作　　者：何延政[1] 张培华[1] 刘建平[2] 王顺德[2] 蒋米尔[1] 陆民[1] 黄新天[1] 林卿[2] 谢毅[2] 

机构地区：[1]上海第二医科大学附属第九人民医院血管外科研究室,200011 [2]复旦大学遗传研究所遗传工程国家重点实验室

出　　处：《中华实验外科杂志》1997年第5期263-264,共2页Chinese Journal of Experimental Surgery

基　　金：上海市科委资助课题

摘　　要：目的:使重组血管内皮细胞生长因子(VEGF)在大肠杆菌中得到高效表达.方法:通过构建表达重组VEGF的质粒PRL621/VEGF,并在大肠杆菌中高效表达.结果:表达量约占菌体总蛋白的40%.对形成包含体的表达产物进行变性,初步复性处理,得到重组人VEGF粗提液,鸡胚绒毛尿囊膜试验表明有促血管生长活性,N-端15个氨基酸序列分析结果,与天然VEGF蛋白质相应序列一致.To study its structure and function, a high level expression plasmid, PRL621/VEGF, which yielded about 30% of total cell protein of Escherichia Coli, was constructed. The inclusion bodies which the uroduction stayed inside were isolated, denatured and then initially renatured. Chorioallantoic membrane (CAM) assay showed the renatured VEGF promoted new vessel formation. Its first 15 N-terminal amino acid sequence was identical to that of the natural protein.

关 键 词：血管内皮细胞 生长因子 大肠杆菌 VEGF 

分 类 号：R378.21[医药卫生—病原生物学] R392－33[医药卫生—基础医学]