蜱传脑炎病毒E抗原蛋白的原核表达及鉴定  

Expression and Identification of Recombinant Tick-Borne Encephalitis Virus Envelope Protein in Escherichia coli

作　　者：温中华[1] 李建民[1] 陈薇[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2017年第4期495-498,共4页Letters in Biotechnology

基　　金："重大新药创制"科技重大专项(2013ZX09304103)

摘　　要：目的:构建蜱传脑炎病毒(TBEV)结构蛋白E的原核表达载体,表达重组蛋白。方法:PCR扩增E蛋白全长基因片段,插入原核表达载体pET-32a并转化大肠杆菌进行诱导表达,镍柱纯化、复性。结果:E蛋白在大肠杆菌中获得表达,表达量达60 mg/L;重组E蛋白能与人抗TBEV多克隆抗体产生特异反应;用重组E抗原免疫家兔后,能检测到较高的抗体水平。结论:原核表达的E抗原蛋白具有抗体结合活性,可用于制备ELISA诊断试剂。Objective： To construct prokaryotic expression vectors for tick-borne encephalitis virus（TBEV） enve- lope（E） protein, and express recombinant TBEV E protein in Escherichia coli. Methods： The coding sequence of full length E protein was amplified by PCR and cloned into the pET-32a vector. The protein of interest was ex- pressed in E.coli with IVFG induction and purified by Ni-NATB bead. Results： We obtained recombinant E pro- tein in E.coli with the yields of -60 mg/L, and it was identified by specific human anti-TBEV polyclonal antibod- ies. After being immunized with E antigen in rabbits, specific high-level antibodies could be detected in serum. Conclusion： The prokaryotically expressed recombinant E protein with the antibody binding activity can be devel- oped as a ELISA diagnostic reagents.

关 键 词：蜱传脑炎病毒 E蛋白 原核表达 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]