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[利用免疫电子显微镜对人41型腺病毒(Ad41)的纤维状包涵体(FIB)进行表征]

[Characterization of fibrillous inclusion body (FIB) of human adenovirus type 41 (Ad41) using immunoelectron microscopy].

作者信息

Song Jing-Dong, Zou Xiao-Hui, Wang Min, Qu Jian-Guo, Lu Zhuo-Zhuang, Hong Tao

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.

出版信息

Bing Du Xue Bao. 2013 Nov;29(6):596-601.

Abstract

To investigate the components of fibrillous inclusion body (FIB), which was formed in packaging cells during the replication of human adenovirus type 41 (Ad41), Ad41 long fiber knob (LFK) and short fiber knob (SFK) proteins were expressed in prokaryote respectively and then used to immunize BALI mice for preparation of anti-LFK serum and anti-SFK sera. The activity and specificity of anti-LFK and an ti-SFK sera were confirmed with Western blot, indirect immunofluorescence assay (IFA) and immunonegative staining, suggesting these sera could be applied in immuno-colloidal gold labelling electron microscopy (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 FIB.

摘要

为研究人41型腺病毒(Ad41)复制过程中在包装细胞内形成的纤维状包涵体(FIB)的成分,分别在原核生物中表达Ad41长纤维钮(LFK)和短纤维钮(SFK)蛋白,然后用于免疫BALB/c小鼠以制备抗LFK血清和抗SFK血清。通过蛋白质免疫印迹法、间接免疫荧光法(IFA)和免疫阴性染色确认抗LFK和抗SFK血清的活性和特异性,表明这些血清可应用于免疫胶体金标记电子显微镜(EM)。用野生型Ad41感染293TE细胞。制作感染细胞的超薄切片,分别用抗Ad41血清、抗LFK血清、抗SFK血清或抗纤维单克隆抗体4D2通过免疫胶体金技术进行标记。在电子显微镜下观察标记的切片,结果表明FIB中同时包含Ad41长纤维蛋白和短纤维蛋白。

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