羊口疮病毒F1L融合Fe蛋白的表达与鉴定  

Expression and identification of orf virus F1L fused with Fe protein

作　　者：邢雪 王元红 李传峰[2] 缪秋红[2] 曹昳 王桂军[1] 刘光清[2] XING Xue;WANG Yuan-hong;LI Chuan-feng;MIAO Qiu-hong;CAO Yi;WANG Gui-jun;LIU Guang-qing(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China;Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Shanghai 200241,China)

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《江苏农业学报》2020年第1期130-135,共6页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2016YFD0500108、2016YFD0501003);上海市科技兴农创新项目[沪农科创字(2019)第3-3号];中央级公益性科研院所基本科研业务费专项(2019JB06)

摘　　要：本研究克隆了羊口疮病毒安徽分离株的F1L基因,融合Fe蛋白编码基因后,插入载体pET-32a中,构建了重组质粒pET-F1L-Fe。将pET-F1L-Fe转化BL21(DE3)感受态细胞,用IPTG诱导表达重组蛋白F1L-Fe。SDS-PAGE分析结果显示,羊口疮病毒F1L-Fe基因在BL21(DE3)获得了正确表达。将大量表达的F1L-Fe蛋白进行纯化,然后免疫BALB/c鼠,制备了抗F1L蛋白的多克隆抗体。最后,以制备的多克隆抗体对F1L-Fe融合蛋白进行Western-blot检测和分析,结果表明F1L-Fe蛋白能与制备的多克隆抗体发生特异性反应,具有良好的反应原性。本研究结果为进一步研发羊口疮病毒亚单位疫苗提供了物质基础。In this study,the F1L gene of orf virus(ORFV)Anhui isolated strain was cloned.After fusing with the coding gene of Fe protein,it was inserted into the expression vector pET-32a to construct recombinant plasmid pET-F1L-Fe.Subsequently,pET-F1L-Fe was transformed into BL21(DE3)competent cells,and the expression of recombinant protein F1L-Fe was induced by IPTG.The results of SDS-PAGE showed that the F1L-Fe gene was successfully expressed in BL21(DE3).Then the F1L-Fe protein was expressed in large quantities and purified.To prepare the polyclonal antibody against F1L protein,BALB/c mice were immunized with the recombinant protein.Western blot results indicated that the F1L-Fe protein reacted specifically with the prepared polyclonal antibody and showed good reactogenicity.In a word,the results of this study provide the material foundation for further developing the subunit vaccine against ORFV.

关 键 词：羊口疮病毒 F1L-Fe蛋白 多克隆抗体 

分 类 号：S855.3[农业科学—临床兽医学]