刺桐胰蛋白酶抑制剂基因的构建及表达研究

Studies on Construction and Expression of Erythrina Trypsin Inhibitor Gene

刺桐胰蛋白酶抑制剂是胰蛋白酶、组织型纤溶酶原激活剂及瑞替普酶特异性的抑制剂 ,可作为配基制成亲合填料用于纯化上述酶。通过化学合成和PCR结合法合成了ETI基因 ,经过序列分析后将其克隆到大肠杆菌表达载体质粒pET32a中 ,并在大肠杆菌中进行诱导表达。表达蛋白分子量同理论值一致 ,生物活性检测表明复性后的ETI对胰蛋白酶及瑞替普酶具有特异抑制作用。

Erythrina Trypsin Inhibitor(ETI) has the ability to bind to trypsin,t-PA and r-PA.So it can be used as an effective ligand for the affinity purification of these enzymes.The ETI gene was synthesised and constructed into the plasmid vector pET32a.The host bacteria containing the recombinant plasmid was induced to express.After crudely purification and renaturation,the fusion protein was analyzed by SDS-PAGE and activity.Molecular mass of the protein are the same with those in theory,and after renaturated the protein can specially bind to trypsin or reteplase.