大肠埃希菌外膜囊泡递呈猪瘟病毒E2蛋白的表达及其免疫效果评价  

Expression of classical swine fever virus E2 protein in outer membrane vesicles of Escherichia coli and evaluation of its immunogenicity

作　　者：黄杰[1] 龚永平[1] 文继峰[1] 杨锐[2] 任露[2] 颜其贵[1,2] HUANG Jie;GONG Yongping;WEN Jifeng;YANG Rui;REN Lu;YAN Qigui(College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;Key Laboratory of Animal Disease and Human Health of Sichuan Province,Chengdu 611130,China)

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]四川农业大学动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《浙江农业学报》2018年第9期1460-1466,共7页Acta Agriculturae Zhejiangensis

基　　金：教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848)

摘　　要：为探讨大肠埃希菌外膜囊泡(outer membrane vesicles,OMVs)作为猪瘟病毒(classical swine fever virus,CSFV)E2蛋白的递呈载体所产生的免疫效应,将切除跨膜区后的CSFV E2基因连接到p BAD18-Cm-Cly A的C-端,转化E.coli DH5α,用0.2%阿拉伯糖诱导表达后,收集含E2蛋白的OMVs免疫家兔。免疫印迹表明,该OMVs可特异性地与组氨酸标签单抗结合。间接ELISA结果表明,重组OMVs可诱导兔体产生高水平的Ig G抗体。免疫后的家兔能很好地防御高剂量CSFV活疫苗攻击。说明大肠埃希菌OMVs递呈CSFV E2蛋白具有较好的免疫原性,为研究OMVs递呈CSFV E2蛋白疫苗奠定了基础。In order to investigate the immune effect of E.coli outer membrane vesicles(OMVs)as presentation vector of classical swine fever virus E2 protein,the classical swine fever virus E 2 gene was ligated to the C-terminus of pBAD18-Cm-Cly A after excision of the transmembrane region,and then transformed into E.coli DH5α.Inducted with 0.2%arabinose,the OMVs containing the E2 protein were collected and the rabbits were immunized.Western blotting showed that the OMVs could specifically bind to CSFV antibody.Indirect ELISA showed that the recombinant OMVs could induce high level of IgG antibody in rabbits.After immunization,rabbits could well protect the live CSFV challenge.All these results indicated that E.coli CSFV E2 protein had good immunogenicity,which laid the foundation for the research of OMVs presentation of classical swine fever virus E2 protein vaccine.

关 键 词：OMVs 猪瘟病毒E2蛋白 免疫原性 

分 类 号：S858.28[农业科学—临床兽医学]