人纤溶酶原饼环区5基因的原核表达及活性测定  

Expression and Its Activity of hPgnK5 Protein in Prokaryotic System

作　　者：范彬[1] 张英起[2] 颜真[2] 赵宁[2] 陈长生[3] 药立波[4] 苏成芝[4] 

机构地区：[1]第四军医大学医学遗传学与发育生物学教研室,西安710032 [2]第四军医大学生物技术中心,西安710032 [3]第四军医大学卫生统计学教研室,西安710032 [4]第四军医大学生物化学教研室,西安710032

出　　处：《中国生物化学与分子生物学报》2001年第6期710-714,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金重点项目 (No .3990 0 12 6 )~~

摘　　要：人纤溶酶原饼环区 5 (hPgnK5 )是新的血管生成抑制因子 .PCR法改造hPgnK5基因 ,所得hPgnK5基因包括编码人纤溶酶原C4 62 到P54 4共 83个氨基酸残基 ,而且在 5′ ,3′分别引入了EcoRⅠ和BamHⅠ位点 .以pThiohisA构建hPgnK5基因原核表达载体 ,在大肠杆菌TOP10中表达该蛋白 .通过柱层析法获得hPgnK5纯化蛋白 .免疫印迹反应表明该蛋白具有hPgnK5免疫活性 .体外实验表明 ,该蛋白具有抑制血管内皮细胞增殖活性 .The human plasminogen kringle 5 is a novel angiogenesis inhibitor.The hPgnK5 gene encoding 83 amino acid residues from C 462 to P 544 was obtained. Eco RⅠ and Bam HⅠ cleavage sites were added to its 5′ and 3′ termini by PCR.The constructed pThiohisA expression vector expressed the hPgnK5 protein in E.coli TOP10.The hPgnK5 protein was purified by column chomatography.Western blot analysis showed that hPgnK5 protein was recognized by mAb against native hPgnK5 and inhibited the proliferation of endothelial cell in vitro. The result shows that the hPgnK5 may be promising in application.

关 键 词：人纤溶酶原饼环区5 血管生成抑制因子 生物活性 基因表达 

分 类 号：Q78[生物学—分子生物学] R394.8[医药卫生—医学遗传学]