马动脉炎病毒GL、M和N蛋白主要抗原域的原核表达及抗原性鉴定  

Prokaryotic expression of the major antigenic domain of equine arteritis GL,M and N protein and antigenicity of their expressed product

作　　者：王贵宾[1,2] 郭巍[1] 赵立平[1] 李红梅[1] 赵敏[2] 相文华[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [2]东北林业大学生命科学学院,黑龙江哈尔滨150040

出　　处：《中国兽医学报》2012年第7期958-961,共4页Chinese Journal of Veterinary Science

基　　金：中央级公益性科研院所基本科研业务费专项(ZGK201007)

摘　　要：采用RT-PCR方法从EAV Bucyrus株扩增了截短的GL、M和全长的N基因片段,将三者分别克隆到表达载体pGEX-6p-1中,测序验证后转化Rosetta(DE3)宿主菌中经IPTG诱导表达,诱导后菌体裂解物经SDS-PAGE分析。试验结果表明,重组菌表达出约35 000、34 000和38 000的特异性条带,通过条件优化及切胶纯化获得较高浓度的目的蛋白。经Western-blot和间接ELISA分析,纯化的蛋白只与抗马动脉炎病毒阳性血清发生特异性反应,证实该蛋白具有良好的反应原性和特异性。The gene of truncated GL and M and the nucleocapsid protein gene were amplified from equine arteritis vi- rus（EAV） Bucyrus strain by RT-PCR and coloned into the pGEX-6p-1 expression vector. After sequencing,the re- combinant plasmid were transformed into Rosetta（DE3）. The transformed bacteria were induced by IPTG and pro- duced a recombinant protein of 35 000,34 000 and 38 000 after purified. The purified protein interacted well with the positive serum against EAV in a Western-blot and indirect ELISA. There results showed that the recombinant pro- tein had good antigenicity and speificity.

关 键 词：马动脉炎病毒 原核表达 抗原性 

分 类 号：S852.65[农业科学—基础兽医学]