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Open AccessArticle

Development of a Honey Bee RNA Virus Vector Based on the Genome of a Deformed Wing Virus

1
USDA, Agricultural Research Service, Bee Research Lab, Beltsville, MD 20705, USA
2
Department of Entomology, University of Maryland, College Park, MD 20742, USA
3
USDA, Agricultural Research Service, Invasive Insect Biocontrol and Behavior Laboratory, Beltsville, MD 20705, USA
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(4), 374; https://doi.org/10.3390/v12040374
Received: 20 February 2020 / Revised: 24 March 2020 / Accepted: 26 March 2020 / Published: 28 March 2020
(This article belongs to the Special Issue Advances in Honey Bee Virus Research)
We developed a honey bee RNA-virus vector based on the genome of a picorna-like Deformed wing virus (DWV), the main viral pathogen of the honey bee (Apis mellifera). To test the potential of DWV to be utilized as a vector, the 717 nt sequence coding for the enhanced green fluorescent protein (eGFP), flanked by the peptides targeted by viral protease, was inserted into an infectious cDNA clone of DWV in-frame between the leader protein and the virus structural protein VP2 genes. The in vitro RNA transcripts from egfp-tagged DWV cDNA clones were infectious when injected into honey bee pupae. Stable DWV particles containing genomic RNA of the recovered DWV with egfp inserts were produced, as evidenced by cesium chloride density gradient centrifugation. These particles were infectious to honey bee pupae when injected intra-abdominally. Fluorescent microscopy showed GFP expression in the infected cells and Western blot analysis demonstrated accumulation of free eGFP rather than its fusions with DWV leader protein (LP) and/or viral protein (VP) 2. Analysis of the progeny egfp-tagged DWV showed gradual accumulation of genome deletions for egfp, providing estimates for the rate of loss of a non-essential gene an insect RNA virus genome during natural infection. View Full-Text
Keywords: honey bee; deformed wing virus; RNA virus vector; invertebrate virus; virus evolution; pollination; food security honey bee; deformed wing virus; RNA virus vector; invertebrate virus; virus evolution; pollination; food security
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MDPI and ACS Style

Ryabov, E.V.; Christmon, K.; Heerman, M.C.; Posada-Florez, F.; Harrison, R.L.; Chen, Y.; Evans, J.D. Development of a Honey Bee RNA Virus Vector Based on the Genome of a Deformed Wing Virus. Viruses 2020, 12, 374.

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