含凝血酶切点的hTNFα-hPgnK_5的原核表达及活性鉴定  

Expression and characterization of fusion protein hTNFα-hPgnK_5 containing thrombin cleavage site in E. coli

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

机构地区：[1]第四军医大学生物学教研室 [2]第四军医大学生物技术中心 [3]第四军医大学生物化学教研室

出　　处：《细胞与分子免疫学杂志》2001年第1期29-31,共3页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的在原核细胞中表达融合蛋白ｈＴＮＦα－ｈＰｇｎＫ５并鉴定其活性。方法构建ｈＴＮＦα－ｈＰｇｎＫ５基因的表达载体，并在大肠杆菌ＤＨ５α中进行表达。以ＭＴＴ法测定该融合蛋白体外抑制肿瘤细胞或血管内皮细胞增殖的活性。结果成功地构建了ｈＴＮＦα－ｈＰｇｎＫ５表达载体并获得表达。免疫印迹分析表明，该融合蛋白具有ｈＰｇｎＫ５的免疫活性。体外实验表明，该融合蛋白具有抑制肿瘤细胞和血管内皮细胞增殖的活性。结论ｈＰｇｎＫ５蛋白可与ｈＴＮＦα可共同表达，表达产物可抑制肿瘤细胞和血管内皮细增殖。Aim To express the fusion protein hTNFα-hPgnK5 in E. Coli and to identify its activity. Methods The hTNFα-hPgnK5 gene express vector was constructed and the fusion protein was expressed in E. coli. The activity of fusion protein to inhibit the proliferation of tumor cells or angioendothelial cells in vitro was determined by MTT. Results Fusion protein hTNFαhPgnK5 possessed immunoactivity in Western blot analysis and could inhibit proliferation of tumor cells or angioendothelial cells in vitro test. Conclusion The hTNFα gene and hPgnk5 gene can be coexpressed in prokaryotic system and expression products can inhibit the proliferation of tumor cells or angioendothelial cells in vitro.

关 键 词：肿瘤坏死因子 血管生长抑制因子 凝血酶切点 hTNFα-hPgnK5 

分 类 号：Q786[生物学—分子生物学] R730.2[医药卫生—肿瘤]