【摘要】 目的 :为了实现蚓激酶基因片段F 3,F 5在真核细胞COS 7中的表达 ,获得具有生物学活性的重组蚓激酶蛋白。方法 :将F 3,F 5与分泌表达载体pcDNA3.1(+)连接 ,构建重组表达质粒pcDNA 3和pcDNA 5 ,利用COS 7细胞进行基因F 3和F 5的瞬时表达 ;纤维蛋白平板法对培养液上清进行活性检测。结果 :蚓激酶基因片段F 3,F 5在真核细胞COS 7中实现了表达 ;SDS PAGE显示蛋白质条带 ,纤维蛋白平板法检测培养液上清有明显的纤溶活性。结论 :蚓激酶基因片段F 3,F 5可在真核细胞COS 7中分泌表达 ,表达产物具有一定的纤溶活性。更多还原

【Abstract】 Objective: To study the biological activity of proteins expressed by lumbrokinase genes in eukaryotic cell COS-7. Methods: The F-3 and F-5 genes were inserted into the eukaryotic expression vector pcDNA3.1(+),respectively. The recombinant plasmids pcDNA-3 and pcDNA-5 were transfected into COS-7 cells. After transfection, the supernatant of culture medium were collected separately at 24 h, 36 h, and 48 h. Using fibrin plate, the fibrinolytic activity of the culture medium was examined. Results: The recombinant expression vector of pcDNA-3 and pcDNA-5 were constructed, and expressed successfully in COS-7 cells. The product in the supernatant of culture medium had obvious fibrinolytic activity defected by fibrin plate method. Conclusion: The genes that encode the F-3 and F-5 can be expressed in the eukaryotic cell COS-7 and the product had the fibrinolytic activity.更多还原