金黄色葡萄球菌B型肠毒素受体拮抗剂的原核表达、纯化及活性初步分析  

Prokaryotic expression,purification and activity analysis of receptor antagonists of staphylococcal enterotoxin B

作　　者：吕艳[1] 王旸[2] 郑玉玲[1] 江华[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [2]北京生物医药研究所,北京100091

出　　处：《现代生物医学进展》2010年第10期1839-1841,共3页Progress in Modern Biomedicine

摘　　要：目的:利用大肠杆菌表达葡萄球菌肠毒素B(SEB)的受体拮抗剂蛋白,检测其与SEB的结合能力,为今后利用其进行SEB快速检测奠定基础。方法:首先确定SEB受体拮抗剂的基因序列,然后将SEB受体拮抗剂质粒转化大肠杆菌BL21(DE3),利用IPTG诱导表达获得蛋白,产物经镍柱纯化后,ELISA鉴定其与SEB结合能力。结果:该质粒在大肠杆菌中获得表达,表达产物可与SEB特异性结合,两者结合能力可达ng/mL。结论:本研究成功对SEB受体拮抗剂进行了原核表达、纯化及初步活性分析。Objective：Receptor antagonists of staphylococcal enterotoxin B were expressed in E.coli,the binding affinity with SEB was determined by ELISA,with aim to facilitate rapid diagnosis of staphylococcal enterotoxin B.Methods：Firstly,the sequence of gene G5-8 was confirmed with the sequence reported,and then the proteins were expressed in E.coli.BL21（DE3） induced by IPTG.The protein was purified by Ni2＋ affinity chromatography,then the binding affinity of the target protein with SEB was detected by ELISA.Results：The protein was expressed successfully in the form of inclusion bodies,and could bind with SEB.with high affinity.Conclusions：The receptor antagonist of staphylococcal enterotoxin B was successfully expressed in prokaryotic expression system.

关 键 词：SEB受体拮抗剂 原核表达 ELISA 

分 类 号：Q939.93[生物学—微生物学] Q75