猪瘟病毒石门株E2基因在永生化猪血管内皮细胞的表达  

Expressing E2 gene of classical swine fever virus Shimen strain in the vein endothelial cells of immortality swine

作　　者：解林红[1] 郭抗抗[1] 张彦明[1] 徐钰[2] 徐彦召[1] 王文秀[1] 

机构地区：[1]西北农林科技大学动物医学学院,陕西杨凌712100 [2]国家林业局森林病虫害防治总站,辽宁沈阳110034

出　　处：《中国预防兽医学报》2008年第6期411-414,419,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金项目(30471290)

摘　　要：利用DNA重组技术将猪瘟病毒石门株囊膜蛋白E2基因定向插入逆转录病毒载体pBABE-puro中,构建重组逆转录病毒载体pBABE-puro-E2。将pBABE-puro-E2和辅助质粒pVSV-G经磷酸钙共沉淀法转染293GP包装细胞系,将其包装为完整的逆转录病毒假病毒;用包装好的假病毒在polybrene的介导下转导第30代的永生化猪脐静脉血管内皮细胞(SUVEC),用嘌呤霉素筛选阳性细胞,并将筛选出的阳性细胞腹腔免疫BALB/c小鼠。RT-PCR检测SUVEC内的E2基因,用间接免疫荧光试验鉴定E2蛋白在SUVEC上的表达,间接ELISA试验检测小鼠血清抗体。结果表明,CSFVE2基因导入猪脐静脉血管内皮细胞获得表达,而且成功诱导小鼠产生抗E2蛋白的抗体。The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting classical swine fever virus （CSFV） Shimen strain E2 gene into pBABE-puro, The pBABE-puro-E2 vector was co-transfected with helper pVSV-G into 293GP packaging cells by calcium phosphate transfection method to produce pseudovirus, which was then used to infect the 30th Swine umbilical vein endothelial cells （SUVEC）, positive cells were screened by puromycine. Expressed E2 protein was detected by RT-PCR and indirect immunofluorescence assays. BALB/c mice were intraperitonally injected with SUVEC expressing E2 protein. Anti-CSFV E2 antibody was detected by ELISA. The results indicated that E2 gene has been successfully expressed in 30th SUVEC induced specific anti-E2 antibody in mouse sera.

关 键 词：猪瘟病毒 E2基因 永生化猪血管内皮细胞 表达 

分 类 号：S852.651[农业科学—基础兽医学]