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  

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

作　　者：姜建[1] 郑玉玲[2] 江华[2] 杨革[1] 

机构地区：[1]曲阜师范大学生命科学学院,山东曲阜273165 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《黔南民族师范学院学报》2012年第3期103-107,69,共6页Journal of Qiannan Normal University for Nationalities

基　　金：国家自然科学基金资助项目(30870091)

摘　　要：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。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 ex- pression vector - PGEX - 4T - 1, and the proteins were expressed in E. coll. BL21 （DE3） induced by IPTG. The protein was purified by GST af- finity chromatography ; the binding affinity of the target protein with SEB was detected by ELISA. The Pharmacedynamics 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： Re- ceptor antagonist of SEB was expressed successfully in soluble form.

关 键 词：金黄色葡萄球菌B型肠毒素受体拮抗剂 原核可溶性表达 ELISA 

分 类 号：R37[医药卫生—病原生物学]