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在尿激酶原cDNA分子对应于Kringle结构域的突环区域 ,插入编码特异四肽Lys-Gly -Asp -Trp(KGDW )分子的DNA片段 ,形成嵌合KGDW序列的尿激酶原突变体全长基因 ,并将其克隆至昆虫细胞杆状病毒表达载体 ,转染昆虫细胞SF9,表达嵌合突变基因 ,获得高效表达 ,对表达产物进行分离纯化获得纯品蛋白。纤维蛋白平板法检测表明嵌合突变体蛋白质具有明显的纤溶酶原激活剂活性。血小板聚集抑制实验表明突变体蛋白质具有明显的血小板聚集抑制活性。该项研究为研制新型溶栓药物打下了基础 ,有望发展成为新型溶栓制剂。
A DNA fragment encoding the specific tetrapeptide Lys-Gly-Asp-Trp (KGDW) molecule was inserted into the prominence region of the urokinase cDNA molecule corresponding to the Kringle domain to form a full length pro-urokinase mutant chimeric KGDW gene , Cloned into insect cell baculovirus expression vector, transfected insect cells SF9, chimeric mutant gene expression, high expression, the expression product was isolated and purified to obtain pure protein. Fibrin plate assay showed that the chimeric mutant protein had significant plasminogen activator activity. Platelet aggregation inhibition experiments showed that the mutant protein has significant platelet aggregation inhibitory activity. The study laid the foundation for the development of new thrombolytic drugs, is expected to develop into a new type of thrombolytic agents.