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Fluorophore labelled BVDV: a novel tool for the analysis of infection dynamics
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Fluorophore labelled BVDV: a novel tool for the analysis of infection dynamics
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Journal Article
Fluorophore labelled BVDV: a novel tool for the analysis of infection dynamics
2019
Overview
Genetic labelling of viruses with a fluorophore allows to study their life cycle in real time, without the need for fixation or staining techniques. Within the family
Flaviviridae
, options for genetic labelling of non-structural proteins exist. Yet, no system to genetically label structural proteins has been put forward to date. Taking advantage of a previously described site within the structural protein E2, a fluorophore was introduced into a cytopathogenic (cpe) BVDV-1 virus (BVDV
E2_fluo
). This insertion was well tolerated, resulting in a 2-fold drop in titer compared to the parental virus, and remained stably integrated into the genome for more than 10 passages. The fluorophore E2 fusion protein was readily detectable in purified virus particles by Western blot and fluorescence microscopy and the particle integrity and morphology was confirmed by cryo electron microscopy. The same integration site could also be used to label the related Classical swine fever virus. Also, BVDV
E2_fluo
particles bound to fluorophore labelled CD46 expressing cells could be resolved in fluorescence microscopy. This underlines the applicability of BVDV
E2_fluo
as a tool to study the dynamics of the whole life cycle of BVDV in real time.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 14/28
/ Animals
/ Bovine Virus Diarrhea-Mucosal Disease - metabolism
/ Bovine Virus Diarrhea-Mucosal Disease - pathology
/ Bovine Virus Diarrhea-Mucosal Disease - virology
/ Cattle
/ Classical Swine Fever - metabolism
/ Classical Swine Fever - pathology
/ Classical Swine Fever - virology
/ Classical Swine Fever Virus - genetics
/ Classical Swine Fever Virus - metabolism
/ Diarrhea Viruses, Bovine Viral - genetics
/ Diarrhea Viruses, Bovine Viral - metabolism
/ Fever
/ Genomes
/ Humanities and Social Sciences
/ Labeling
/ Membrane Cofactor Protein - metabolism
/ Microscopy, Fluorescence - methods
/ Science
/ Swine
/ Viral Envelope Proteins - genetics
/ Viral Envelope Proteins - metabolism
/ Viruses
