Production of a hybrid protein consisting of the N-terminal fragment of urokinase and the C-terminal domain of urinary trypsin inhibitor in Escherichia coli.
Production of a hybrid protein consisting of the N-terminal fragment of urokinase and the C-terminal domain of urinary trypsin inhibitor in Escherichia coli.
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在大肠杆菌中生产由尿激酶 N 端片段和尿胰蛋白酶抑制剂 C 端结构域组成的杂合蛋白。
DOI：
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发表时间：
1998
影响因子：
2.8
通讯作者：
T. Terao
中科院分区：
文献类型：
--
作者：
D. Sugino;M. Okushima;H. Kobayashi;T. Terao
我们构建了一个由尿激酶（ATF）的N端片段和尿胰蛋白酶抑制剂的C端结构域HI-8组成的杂合蛋白（ATFHI）。通过PCR工程化杂合蛋白的融合基因并克隆到表达质粒中。在tac启动子的调控下，融合基因在大肠杆菌中得到了高效表达。在大肠杆菌中以包涵体形式表达的杂合蛋白。大肠杆菌，通过透析方法复性，并通过离子交换层析纯化。ATFHI具有与抗转移作用相关的双功能活性：ATF的尿激酶受体结合活性和HI-8对纤溶酶的抑制活性。
We have constructed a hybrid protein (ATFHI) consisting of an N-terminal fragment from urokinase (ATF) and HI-8, which is the C-terminal domain of urinary trypsin inhibitor. The fusion genes for the hybrid proteins were engineered by PCR and cloned into expression plasmids. Under the control of the tac promoter, fusion genes were efficiently expressed in Escherichia coli. The hybrid proteins, produced as inclusion bodies in E. coli, were refolded by a dialysis method and purified by ion-exchange chromatography. ATFHI exhibited bifunctional activity related to antimetastatic effects: the urokinase receptor-binding activity of ATF and the inhibitory activity of HI-8 on plasmin.