猪瘟病毒E2基因密码子优化后在昆虫细胞中的表达  

The optimization of CSFV E2 gene condons and expression by insect cells

作　　者：范学政[1] 徐璐[1] 王琴[1] 赵启祖[1] 宁宜宝[1] 陈锴[1] 

机构地区：[1]中国兽医药品监察所猪瘟参考实验室,北京海淀100081

出　　处：《中国兽医杂志》2011年第5期3-5,共3页Chinese Journal of Veterinary Medicine

基　　金：农业部"948"项目(2006-G57);国家科技支撑计划(2006BAD06A12)

摘　　要：将猪瘟病毒E2基因密码子改造后,克隆入pFastBac1质粒载体,经转座、转染后构建重组杆状病毒。并对表达产物进行鉴定分析。结果:E2基因改造后与设计序列一致;成功获得了重组杆状病毒(BAC/SE2M),采用猪瘟E2单抗WH303进行Western-blot和ELISA鉴定,证明:表达的CSFV E2蛋白特异性好,纯化后的E2蛋白,可用于猪瘟抗体检测试剂盒的开发。The E2 gene of classical swine fever virus（CSFV） was optimized in order to be expressed in baculovirus.The modified E2 gene of CSFV,honeybee melittin signal（Mels） sequence and six-histidine tag were fused and cloned into vector pFastBac1.After transformation to DH10Bac and transfection to sf9,a recombinant virus named BAC/SE2M was constructed.The supernatant was concentrated and purified with Nie＋ affinity chromatography.Purified E2 protein can be recognized by mAb WH303.Recombinant E2 protein expressed by insect cells can be used to develop ELISA for CSFV Ab.

关 键 词：猪瘟病毒 E2基因 密码子优化 杆状病毒 

分 类 号：S852.651[农业科学—基础兽医学]