含马动脉炎病毒N基因的病毒样颗粒的制备  

Construction of virus-like particles containing N gene of equine arteritis virus

作　　者：王宇[1,2] 张晓文[2] 王群[2] 梁成珠[2] 汪东风[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛260003 [2]山东出入境检验检疫局检验检疫技术中心,山东青岛266002

出　　处：《畜牧与兽医》2014年第6期8-11,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家质检总局科技计划项目(2011IK010)

摘　　要：为了构建易于纯化且耐RNase酶的含有马动脉炎病毒N基因的病毒样颗粒,通过RT-RCR扩增马病毒性动脉炎病毒的N基因,将其克隆至含有组氨酸标签的pNH-MS2his载体中,构建重组原核表达载体pNH-MS2his-N。将重组质粒pNH-MS2his-N转化表达菌株大肠杆菌BL21(DE3),1 mmol/L IPTG诱导表达、纯化。对病毒样颗粒进行鉴定及稳定性实验。结果表明该病毒样颗粒含马病毒性动脉炎病毒N基因,并且稳定性良好,构建的病毒样颗粒可以作为RNA病毒检测时的标准品和质控品使用。The aim of this study is to construct purifyable and RNase-resistant virus-like particles （VLPs） containing N gene of equine arteritis virus （EAV）. The N gene of EAV was amplified by RT-PCR, then the gene was cloned into vector pNH-MS2his with MS2 phage coat protein, mature enzyme gene and histidine-tag to construct the prokaryotic expression vector pNH-MS2his-N. The recombinant plasmid pNH-MS2his-N was transformed into E. coli strain BL21 （DE3） and induced to express with 1 mmol/L IPTG. The virus-like particles （VLPs） were identified and the stability of VLPs was tested by real-time RT-PCR. The results showed that the VLPs contain N gene and have good stability. The VLPs can be used as a quality control in RNA virus detection.

关 键 词：马动脉炎病毒 病毒样颗粒 装甲RNA 

分 类 号：S852.652[农业科学—基础兽医学]