血管生成抑制素K1～3环结构体酵母表达及活性检测

Expression of kringlie 1～3 of angiostatin in Pichia and activity detection

目的研究血管生成抑制素K1～ 3重组体的生物学活性及机理。方法将血管生成抑制素K1～ 3环结构基因克隆进入甲醇表达质粒 pPIC9K中 ,诱导该酵母表达系统表达了K1～ 3重组蛋白 ,经初步纯化后 ,利用改良MTT法在人脐静脉内皮细胞基质上检测其对血管内皮细胞的生长抑制活性。结果确认血管生成抑制素K1～ 3重组体能明显抑制人脐静脉内皮细胞的生长。结论血管生成抑制素K1～ 3重组体抑制能力具有量 效关系 ,抑制该内皮细胞生长的基本机理是诱导其发生凋亡类似反应。

PurposeThe aim is to invest the mechanism and activity of kringle1～3 gene of angiostatin. MethodsKringle1～3 gene of angiostatin was abtained from hepatocyte of normal human by RT PCR and inserted into expression vector pPIC9K in Pichia expression system. The recombinant protein was induced secrete , purified and test the activity by MTT.ResultsKrigle1～3 protein could inhibit vascular endothelial cell proliferative activity.ConclusionThe reaction similar apoptosis induced by kringle1～3 recombinant protein of angiostatin inhibit cell proliferation.