机构地区: 中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室
出 处: 《中国生物工程杂志》 2006年第4期27-31,共5页
摘 要: 目的:构建凝血酶敏感蛋白-1N端片段(TSF)的原核表达体系,测定重组蛋白的活性。方法:从人脐静脉内皮细胞cDNA中扩增得到thbs1基因片段,克隆到pET32c(+)载体系统并转化 BL21大肠杆菌,表达纯化得到TSF。MTT法体外测定目的蛋白对ECV304细胞增殖的抑制作用。免疫荧光标记流式细胞仪测定CD36的表达。结果:获得了原核表达的重组TSF蛋白。目的蛋白对CD36阴性的ECV304细胞有增殖抑制作用。结论:低剂量重组TSF蛋白是一个具有临床应用前景的抗肿瘤治疗的辅助方法。 Objective: To construct an expression system containing the N-terminal segment of Thrombospondin-1 (TSP-1) in E. coli, To investigate the activity of TSF, Methods : thbsl gene fragment was amplified from human fetal cord vein endothelial cells and inserted into plasmid pE332c ( + ) which was then transfected and expressed in E. coli. Investigate the effect of recombinant TSF to the pro.liferation of ECV304 in vitro with MTT. The expression level of CD36 was assayed by FACS, Results: Recombinant TSF was purified, TSF could restrain the proliferation of ECV304 with CD36 low-expression, Conclusions : Low dose of rTSF was a latent assistant treatment of anti-cancer.