人腺病毒41型纤维丝状包涵体免疫电镜研究  

Characterization of Fibrillous Inclusion Body(FIB)of Human Adenovirus Type41(Ad41)Using Immunoelectron Microscopy

作　　者：宋敬东[1] 邹小辉[1] 王敏[1] 屈建国[1] 鲁茁壮[1] 洪涛[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室,北京100052

出　　处：《病毒学报》2013年第6期596-601,共6页Chinese Journal of Virology

基　　金：国家科技重大专项(2011ZX10004-001);中国疾病预防控制中心青年科研基金(2013A105)

摘　　要：为明确人腺病毒41型(Adenovirus type 41,Ad41)形态发生过程中形成的纤维丝状包涵体(Fibrillous inclusion body,FIB)是否含有Ad41的长纤维(Long fiber,LF)蛋白与短纤维(Short fiber,SF)蛋白。我们分别原核表达和纯化Ad41LF、SF的球部(Knob)蛋白,分别命名为LFK与SFK。以LFK和SFK分别免疫BALB/c小鼠,分别获得LFK抗血清和SFK抗血清。Western blot、间接免疫荧光和免疫负染结果表明,LFK抗血清和SFK抗血清分别与LF和SF结合,LFK抗血清和SFK抗血清之间无交叉反应,可以应用于免疫电镜标记。通过Ad41抗血清、腺病毒纤维单克隆抗体4D2、LFK抗血清与SFK抗血清分别对FIB进行免疫电镜胶体金标记,表明Ad41抗血清、腺病毒纤维单克隆抗体4D2、LFK抗血清与SFK抗血清均能标记FIB,说明FIB中含有Ad41长纤维蛋白和短纤维蛋白。To investigate the components of fibrillous inclusion body （FIB）, which was formed in packaging cells during the replication of human adenovirus type 41 fAd41）, Ad41 long fiber knob （LFK） and short fiber knob （SFK） proteins were expressed in prokaryote respectively and then used to immunize BALB/c mice for preparation of anti-LFK serum and antbSFK sera. The activity and specificity of anti-LFK and an- ti-SFK sera were confirmed with Western blot, indirect immunofluorescence assay （IFA） and immuno- negative staining, suggesting these sera could be applied in immuno-colloidal gold labelling electron mi- croscopy （EM）. 293TE cells were infected with wild-type Ad41. Ultrathin sections of infected cells were made, and labelled with immuno-colloidal gold technique using anti-Ad41 sera, anti-LFK sera, anti-SFK sera, or anti-fiber monoclonal antibody 4D2, respectively. The labelled sections were observed under EM, and the results demonstrated that both Ad41 long fiber protein and short fiber protein were included in FIB.

关 键 词：人腺病毒41型 纤维丝状包涵体 免疫电镜技术 

分 类 号：R373.9[医药卫生—病原生物学]