一种用于生产重组尿激酶型纤溶酶原激活剂的高效系统。
An efficient system for production of recombinant urokinase-type plasminogen activator.
作者信息
Tang W, Sun Z Y, Pannell R, Gurewich V, Liu J N
机构信息
Vascular Research Laboratory, Beth Israel-Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
出版信息
Protein Expr Purif. 1997 Dec;11(3):279-83. doi: 10.1006/prep.1997.0800.
A system was developed to produce recombinant urokinase-type plasminogen activator in Escherichia coli. The urokinase-type plasminogen activator was produced with a 6 x His-tag at the C-terminus which was shown to have the same activity, after refolding, as the wild-type protein. Purification of the recombinant protein to homogeneity (95%) was possible by one-step affinity chromatography under denaturing conditions. As a result, proteolysis by bacterial proteases during purification was avoided. A higher refolding efficiency (40%) and a higher total recovery yield (25%) of the recombinant protein were obtained by this method.
已开发出一种在大肠杆菌中生产重组尿激酶型纤溶酶原激活剂的系统。尿激酶型纤溶酶原激活剂在C端带有6×His标签，复性后显示出与野生型蛋白相同的活性。在变性条件下通过一步亲和层析可将重组蛋白纯化至同质（95%）。因此，避免了纯化过程中细菌蛋白酶的蛋白水解作用。通过该方法获得了更高的复性效率（40%）和更高的重组蛋白总回收率（25%）。
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