Intracellular localisation of dengue-2 RNA in mosquito cell culture using electron microscopic in situ hybridisation

Intracellular localisation of dengue-2 RNA in mosquito cell culture using electron microscopic in... Non-isotopic in situ hybridisation was used at the electron microscope level to determine the localisation of viral RNA in dengue-2 infected mosquito cells at 14, 24, 48 and 72 h post-infection. In situ hybridisation was carried out on sections of dengue-2 infected mosquito cells using a digoxigenin-labelled DNA probe to the envelope protein gene sequence of the virus. Viral RNA was consistently localised over the rough endoplasmic reticulum and the virus-induced smooth membrane structures which form within the endoplasmic reticulum. During the later stages of infection electron-dense areas were observed to develop in close proximity to the smooth membrane structures. Electron microscopic in situ hybridisation showed that these denser areas contained both viral RNA and virus particles. Our results show that in dengue-2 infected mosquito cells the smooth membrane structures are an important site for the concentration of dengue viral RNA and its possible subsequent encapsidation into virus particles. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Intracellular localisation of dengue-2 RNA in mosquito cell culture using electron microscopic in situ hybridisation

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Publisher
Springer-Verlag
Copyright
Copyright © Wien by 1997 Springer-Verlag/
Subject
Legacy
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s007050050247
Publisher site
See Article on Publisher Site

Abstract

Non-isotopic in situ hybridisation was used at the electron microscope level to determine the localisation of viral RNA in dengue-2 infected mosquito cells at 14, 24, 48 and 72 h post-infection. In situ hybridisation was carried out on sections of dengue-2 infected mosquito cells using a digoxigenin-labelled DNA probe to the envelope protein gene sequence of the virus. Viral RNA was consistently localised over the rough endoplasmic reticulum and the virus-induced smooth membrane structures which form within the endoplasmic reticulum. During the later stages of infection electron-dense areas were observed to develop in close proximity to the smooth membrane structures. Electron microscopic in situ hybridisation showed that these denser areas contained both viral RNA and virus particles. Our results show that in dengue-2 infected mosquito cells the smooth membrane structures are an important site for the concentration of dengue viral RNA and its possible subsequent encapsidation into virus particles.

Journal

Archives of VirologySpringer Journals

Published: Dec 1, 1997

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