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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 determine whether the Fibrillous inclusion body (FIB) formed during the morphogenesis of human adenovirus type 41 (Ad41) contains the Ad41 long fiber (LF) protein and the short fiber (Short fiber) fiber, SF) protein. We prokaryotic expression and purification of Ad41LF, SF Knob protein, respectively, named LFK and SFK. BALB / c mice were immunized with LFK and SFK respectively to obtain LFK antiserum and SFK antiserum respectively. Western blot, indirect immunofluorescence and immunofluorescence staining showed that there was no cross-reaction between LFK antiserum and SFK antiserum, and LFK antiserum and SFK antiserum could be used for immunoelectron microscopy. Ad41 antiserum, adenovirus fiber monoclonal antibody 4D2, LFK antiserum and SFK antiserum were used for immunoelectron microscopic gold staining of FIB, which indicated that Ad41 antiserum, adenovirus fiber monoclonal antibody 4D2, LFK antiserum and SFK antiserum FIB can be labeled, indicating that FIB contains Ad41 long fibrin and short fibrin.