Angiostatin K(1-3)基因的原核表达和活性鉴定

PROKARYOTIC EXPRESSION AND IDENTIFICATION OF THE GENE FRAGMENT OF ANGIOSTATIN K(1 3)

将人AngiostatinK( 1 3)基因在原核细胞表达并鉴定其活性。构建和鉴定AngiostatinK ( 1 3)原核表达载体并在大肠杆菌DH5α内表达 ,诱导表达蛋白用SDS PAGE分析和Westernblot鉴定 ,采用人脐带静脉内皮细胞抑制实验和鸡胚绒毛膜尿囊膜试验验证其活性。AngiostatinK( 1 3)功能区段基因在原核细胞获得成功表达 ,目的蛋白占菌体总蛋白的 1 5 .7%。该表达蛋白能抑制内皮细胞增殖活性 ,具有剂量依赖效应。正确表达的AngiostatinK( 1 3)基因为实体瘤抗血管生成治疗提供了条件。

To express angiostatinK(1 3) cDNA in E.coli and identify its protein activity.The vector pDH angiostatin K(1 3) was constructed, identified and expressed inE.coli DH5α.After induction and affinity chromatography purification, the protein was analyzed in SDS PAGE and identified by Western blot. Its activity was examined by human funicular vein endotheliocyte inhibition and chicken chorio allantoic membrane test. The human angiostatin K(1 3) gene fragment was sucessfully expressed in Escherichia coli. The target protein made up 15.7%of the total bacterial protein. The angiostatin K(1 3) protein could inhibit proliferation of endothelial cells, andhad a dose dependenteffect.The angiostatinK(1 3) gene, accurately expressed , is of great significance for tumor antiangiogenesis therapy.