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超薄冷冻切片在感染的非洲绿猴肾细胞培养物中定位流感病毒抗原的应用。

The use of ultrathin cryosections for localisation of influenza virus antigens in infected vero cell cultures.

作者信息

Beesley J E, Campbell D A

出版信息

Histochemistry. 1984;80(5):497-501. doi: 10.1007/BF00495441.

DOI:10.1007/BF00495441
PMID:6480415
Abstract

The localisation of influenza virus antigens in infected Vero cell monolayer cultures by post embedding immunoelectron microscopy requires both good resolution and the retention of antigenicity in the tissue sections. Ultrathin cryosections are superior to ultrathin resin sections for this purpose. The colloidal gold probe was used in conjunction with specific antibody preparations to localise three viral proteins. Antibody raised against haemagglutinin glycoproteins labelled the host cell membrane and the virus fringe without contamination of the host cell nucleus, whereas antibody raised against viral nuclear protein labelled throughout the host cell cytoplasm and nucleus. Matrix protein was localised within the nucleus and was associated with the host cell membrane of the infected cell. The appearance of all these proteins was maximal 24 h post infection.

摘要

通过包埋后免疫电子显微镜对感染的非洲绿猴肾细胞单层培养物中的流感病毒抗原进行定位,既需要良好的分辨率,又需要在组织切片中保留抗原性。为此,超薄冷冻切片优于超薄树脂切片。胶体金探针与特异性抗体制剂联合使用,以定位三种病毒蛋白。针对血凝素糖蛋白产生的抗体标记了宿主细胞膜和病毒边缘,而未污染宿主细胞核,而针对病毒核蛋白产生的抗体则在整个宿主细胞质和细胞核中进行标记。基质蛋白定位于细胞核内,并与感染细胞的宿主细胞膜相关。所有这些蛋白在感染后24小时出现的量最大。

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引用本文的文献

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本文引用的文献

1
THE APPLICATION OF FERRITIN-CONJUGATED ANTIBODY TO ELECTRON MICROSCOPIC STUDIES OF INFLUENZA VIRUS IN INFECTED CELLS : I. THE CELLULAR SURFACE.铁蛋白结合抗体在感染细胞中流感病毒电子显微镜研究中的应用:I. 细胞表面。
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Immunochemistry on ultrathin frozen sections.超薄冰冻切片的免疫化学
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Migration of influenza virus-specific polypeptides from cytoplasm to nucleus of infected cells.流感病毒特异性多肽从受感染细胞的细胞质迁移至细胞核。
Virology. 1981 May;111(1):154-64. doi: 10.1016/0042-6822(81)90661-9.
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Influenza virus RNA is synthesized at fixed sites in the nucleus.流感病毒核糖核酸在细胞核内的固定位点合成。
Nature. 1982 Mar 25;296(5855):366-8. doi: 10.1038/296366a0.
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Antibody labeling of thin sections of skeletal muscle with specific antibodies: a comparison of bovine serum albumin (BSA) embedding and ultracryomicrotomy.
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Postembedding immunocytochemical localization of paramyxovirus antigens by light and electron microscopy.通过光学显微镜和电子显微镜对副粘病毒抗原进行包埋后免疫细胞化学定位。
J Histochem Cytochem. 1982 Dec;30(12):1313-9. doi: 10.1177/30.12.6296222.
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Specific antisera produced by direct immunization with slices of polyacrylamide gel containing small amounts of protein.通过用含有少量蛋白质的聚丙烯酰胺凝胶切片直接免疫产生的特异性抗血清。
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Cleavage of structural proteins during the assembly of the head of bacteriophage T4.在噬菌体T4头部组装过程中结构蛋白的切割
Nature. 1970 Aug 15;227(5259):680-5. doi: 10.1038/227680a0.
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Influenza virus-induced proteins in nuclei and cytoplasm of infected cells.流感病毒诱导感染细胞的细胞核和细胞质中的蛋白质。
Virology. 1977 Jun 15;79(2):449-54. doi: 10.1016/0042-6822(77)90372-5.