SEB超抗原受体拮抗剂在大肠杆菌中的高效可溶性表达、纯化及活性初步研究

目的:在大肠杆菌(BL21)中构建可溶性表达的金黄色葡萄球菌B型肠毒素(SEB)受体拮抗剂。方法:首先确定SEB受体桔抗剂的基因序列,然后用含有SEB受体桔抗剂的基因序列重组质粒表达载体PGEX-4T-1转化大肠杆菌BL21(DE3),利用IPTG诱导表达获得蛋白,产物经GST柱纯化后,利用ELISA检测其与SEB的结合能力并进行其体内外药效学实验。结果:该质粒成功转化为可溶性表达,ELISA结果显示表达产物可与SEB特异性结合。结论:本研究成功对SEB受体拮抗剂GST可溶性表达并对其活性进行初步分析。

Soluble High - expression, Purification and Biological Activity Analysis of Receptor Antagonist of SEB in Escherichia coli

Objective:Receptor antagonist of staphylococcal enterotoxin B(SEB) was high-expressed in E.coli(BL21 ).Methods:Firstly, the sequence of gene G5-8 was confirmed with the reported sequence,then the sequence of gene G5-8 was cloned and constructed a plasmid expression vector-PGEX-4T-1,and the proteins were expressed in E.coli.BL21(DE3) induced by IPTG.The protein was purified by GST affinity chromatography;the binding affinity of the target protein with SEB was detected by ELISA.The Pharmacodynamics of the protein in vitro and in vivo was studied.Results:Receptor antagonists of SEB were expressed successfully in soluble form,and could bind with SEB.Conclusions:Receptor antagonist of SEB was expressed successfully in soluble form.