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Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

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Elliott2013JGenVir94_851.pdf (530.1Kb)
Date
04/2013
Author
Elliott, Richard M.
Blakqori, Gjon
van Knippenberg, Ingeborg C.
Koudriakova, Elina
Li, Ping
McLees, Angela
Shi, Xiaohong
Szemiel, Agnieszka M.
Funder
The Wellcome Trust
BBSRC
Medical Research Council
Grant ID
079810/Z/06/Z
BB/G004277/1
G0800161
Keywords
Valley fever virus
Defective interfering particles
CDNA-based rescue
NSS protein
Untranslated regions
Family bunyaviridae
Replication
RNA
Lacking
Genome
QR355 Virology
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Abstract
Schmallenberg virus (SBV) is a newly emerged orthobunyavirus that has caused widespread disease in cattle, sheep and goats in Europe. Like other orthobunyaviruses, SBV is characterized by a tripartite negative-sense RNA genome that encodes four structural and two non-structural proteins. This study showed that SBV has a wide in vitro host range, and that BHK-21 cells are a convenient host for both SBV propagation and assay by plaque titration. The SBV genome segments were cloned as cDNA and a three-plasmid rescue system was established to recover infectious virus. Recombinant virus behaved similarly in cell culture to authentic virus. The ORF for the non-structural NSs protein, encoded on the smallest genome segment, was disrupted by introduction of translation stop codons in the appropriate cDNA, and when this plasmid was used in reverse genetics, a recombinant virus that lacked NSs expression was recovered. This virus had reduced capacity to shut-off host-cell protein synthesis compared with the wild-type virus. In addition, the NSs-deleted virus induced interferon (IFN) in cells, indicating that, like other orthobunyaviruses, NSs functions as an IFN antagonist, most probably by globally inhibiting host-cell metabolism. The development of a robust reverse genetics system for SBV will facilitate investigation of its pathogenic mechanisms as well as the creation of attenuated strains that could be candidate vaccines.
Citation
Elliott , R M , Blakqori , G , van Knippenberg , I C , Koudriakova , E , Li , P , McLees , A , Shi , X & Szemiel , A M 2013 , ' Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe ' , Journal of General Virology , vol. 94 , no. 4 , pp. 851-859 . https://doi.org/10.1099/vir.0.049981-0
Publication
Journal of General Virology
Status
Peer reviewed
DOI
https://doi.org/10.1099/vir.0.049981-0
ISSN
0022-1317
Type
Journal article
Rights
Copyright (c) 2013 SGM. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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  • University of St Andrews Research
URI
http://hdl.handle.net/10023/3908

Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.

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