Tracing Baculovirus AcMNPV Infection Using a Real-Time Method Based on ANCHOR TM DNA Labeling Technology
| Autor: | Linda A. King, Sylvie Juliant, Miguel López Ferber, Aurélie Hinsberger, Benoit Graillot, Christine Blachère Lopez, Robert D. Possee, Franck Gallardo, Martine Cerutti |
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| Přispěvatelé: | Eau, Ressources, Territoires (ERT - IMT Mines Alès), Laboratoire de Génie de l'Environnement Industriel et des Risques Industriels et Naturels (LGEI), IMT - MINES ALES (IMT - MINES ALES), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT)-IMT - MINES ALES (IMT - MINES ALES), Institut Mines-Télécom [Paris] (IMT)-Institut Mines-Télécom [Paris] (IMT), INRA Sophia Antipolis, Institut National de la Recherche Agronomique (INRA), Baculovirus et Thérapie, Centre National de la Recherche Scientifique (CNRS), Department of Biological and Medical Sciences, Oxford Brookes University, ITAV-USR 3505, Université Fédérale Toulouse Midi-Pyrénées, NeoVirTech SAS, This research was funded by the Region Occitanie doctoral grant number 126/2017 for A.H., Institut Sophia Agrobiotech (ISA), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Recherche Agronomique (INRA), Institut des Technologies Avancées en sciences du Vivant (ITAV), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
viruses Green Fluorescent Proteins 030106 microbiology lcsh:QR1-502 Sf9 Genome Viral Spodoptera Virus Replication Genome baculovirus infection lcsh:Microbiology Article Virus 03 medical and health sciences chemistry.chemical_compound [SPI]Engineering Sciences [physics] fluorescence labelling In vivo Virology Sf9 Cells Animals Fluorometry real-time imaging biology digestive oral and skin physiology fungi biology.organism_classification Nucleopolyhedroviruses In vitro 3. Good health DNA-Binding Proteins Autographa californica 030104 developmental biology Infectious Diseases Microscopy Fluorescence chemistry Larva DNA Viral DNA |
| Zdroj: | Viruses Viruses, MDPI, 2020, 12 (1), art.50 (11 p.). ⟨10.3390/v12010050⟩ Viruses, Vol 12, Iss 1, p 50 (2020) Volume 12 Issue 1 |
| ISSN: | 1999-4915 |
| Popis: | Many steps in the baculovirus life cycle, from initial ingestion to the subsequent infection of all larval cells, remain largely unknown primarily because it has hitherto not been possible to follow individual genomes and their lineages. Use of ANCHORTM technology allows a high intensity fluorescent labelling of DNA. When applied to a virus genome, it is possible to follow individual particles, and the overall course of infection. This technology has been adapted to enable labelling of the baculovirus Autographa californica Multiple NucleoPolyhedroVirus genome, as a first step to its application to other baculoviruses. AcMNPV was modified by inserting the two components of ANCHORTM: a specific DNA-binding protein fused to a fluorescent reporter, and the corresponding DNA recognition sequence. The resulting modified virus was stable, infectious, and replicated correctly in Spodoptera frugiperda 9 (Sf9) cells and in vivo. Both budded viruses and occlusion bodies were clearly distinguishable, and infecting cells or larvae allowed the infection process to be monitored in living cells or tissues. The level of fluorescence in the culture medium of infected cells in vitro showed a good correlation with the number of infectious budded viruses. A cassette that can be used in other baculoviruses has been designed. Altogether our results introduce for the first time the generation of autofluorescent baculovirus and their application to follow infection dynamics directly in living cells or tissues. |
| Databáze: | OpenAIRE |
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