重组人纤溶酶原Kringle1-3的生物学活性鉴定

目的：检测重组人纤溶酶原Kringle1-3（K1-3）的生物学活性。方法：用含重组人纤溶酶原K1-3基因的表达载体pET21a-Angio（K1-3）转化表达宿主菌大肠杆菌BL21（DE3）后诱导表达,表达产物经溶解、复性和纯化后,进行SDS-PAGE,计算其相对分子质量;用BCA法测蛋白浓度,用细胞抑制实验（MTT法）和鸡胚绒毛膜尿囊膜（CAM）实验鉴定纯化产物对血管内皮细胞增殖和血管生成的抑制效果。结果：表达产物的相对分子质量为38000,与预期值一致;细胞抑制实验和CAM实验结果表明表达产物具有特异抑制血管内皮细胞增殖、血管生成的功能。结论：所纯化的重组人纤溶酶原K1-3具有抑制血管内皮细胞的生物学活性,为该蛋白在病理性血管疾病等方面的应用研究提供了材料。

Anti-Angiogenesis Assay of Recombinant Kringle1-3 Domain of Human Plasminogen

Objective： To assay biological activity of recombinant human plasminogen Kringlel-3（K1-3）. Methods： The recombinant protein was expressed in E.coli BL21（DE3） and purified by affinity chromatography, and its respective molecular weight （Mr） was determined according to SDS-PAGE figure. After concentration of purified protein was measured by BCA test, the biological activity was examined by endothelial cell restraint test and angiogenesis restraint test of chick ambryonal membrane（CAM）. Results： The Mr of expression product was 38 000 as expected. The recombinant K1-3 exhib- ited the inhibitory activity of endothelial cell proliferation and restraining angiogenesis. Conclusion： The recombinant hu- man plasminogen K1-3 has anti-angiogenesis activity, and showed the potential of clinical application.