Journal Article

Co-localization of HSV-1 DNA and ICP35 protein by <i>in situ</i> hybridization and immunocytochemistry

Hiroyuki Morioka, Kappei Kobayashi, Masayoshi Tachibana and Jiro Imanishi

in Microscopy

Published on behalf of The Japanese Society of Microscopy

Volume 48, issue 5, pages 621-628
Published in print January 1999 | ISSN: 2050-5698
Published online January 1999 | e-ISSN: 2050-5701 | DOI: http://dx.doi.org/10.1093/oxfordjournals.jmicro.a023728
Co-localization of HSV-1 DNA and ICP35 protein by in situ hybridization and immunocytochemistry

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In an effort to obtain a better signal-to-noise ratio and ultrastructural preservation, we sought to improve electron microscopic in situ hybridization technique. In our method, protease treatment was omitted and visualization of the digoxigenin (DIG)-labelled deoxyribonucleic acid (DNA)-probe was enhanced using a three-step procedure. These improvements allowed us to localize viral DNA with good signal-to-noise ratio. DNA specific to herpes simplex virus 1 (HSV-1) was localized by this method to HSV-1 infected cultured cells; DNA was not observed in the empty-cored HSV- 1. Using this method and the immunogold cytochemical method, we co-localized viral DNA and capsid protein ICP35 on Lowicryl-embedded sections of HSV-1 infected cells. Interestingly, labelling for both DNA and ICP was observed on some HSV-1 particles in cell nucleus. This finding is consistent with the notion that ICP35 is necessary for assembly of viral DNA. Combination of in situ hybridization, and immunocytochemical techniques is a powerful tool for examination of the functional relationship between viral DNA and proteins and help us to study protein function in viral multiplication.

Keywords: herpes simplex virus type 1; electron microscopic in situ hybridization; dioxigenin-labelled DNA; electronmicroscopic immunogold cytochemistry; ICP35; IgG-gold

Journal Article.  0 words. 

Subjects: Biological Sciences

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