摘要
以猪瘟病毒石门株E2囊膜糖蛋白基因序列为基础,构建了3种重组杆状病毒rAc-E2、rAc-E2oKm和rAc-HE2oKm。Western-blot和间接免疫荧光试验结果显示,这3种重组杆状病毒均可正确表达重组E2蛋白,与E2蛋白和His的单克隆抗体均有良好的抗原反应特性,其中rAc-HE2oKm可分泌表达重组E2蛋白,可形成病毒样颗粒,可诱导免疫仔猪产生高水平抗体和中和抗体,免疫效果与猪瘟活疫苗相当,为猪瘟病毒E2蛋白亚单位疫苗研究奠定了重要基础。
Based on E2 gene of classical swine fever virus(CSFV) Shimen strain,three recombinant baculoviruses r Ac-E2,r Ac-E2oKm and r Ac-HE2oKm,expressing the modified E2 genes,were constructed and identified by indirect immunofluorescent assay and Western-blot with monoclonal antibodies His-Tag and 7H2.The results showed that the three modified E2 proteins could be expressed in Sf9 cells and react with the monoclonal antibodies efficiently.Among them,r Ac-HE2oKm could secretory express the E2 protein.And the recombinant E2 protein,HE2oKm could assemble as virus-like particles.The piglets immunized with r Ac-HE2oKm developed high-level anti-CSFV antibodies detected by ELISA and neutralizing assay.The immune effect was similar to that of live classical swine fever vaccine,which laid an important foundation for the study of CSFV E2 protein subunit vaccine.
作者
高莹
白娟
宋中宝
姜平
GAO Ying;BAI Juan;SONG Zhong-bao;JIANG Ping(Key Laboratory of Animal Bacteriology,Ministry of Agriculture/College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2019年第8期983-989,共7页
Chinese Veterinary Science
基金
国家生猪产业技术体系专项(CARS-35)
国家重点研发计划项目(2018YFD0500803)
关键词
猪瘟病毒
E2蛋白
亚单位疫苗
classical swine fever virus
E2 protein
subunit vaccine
