dc.contributor.author |
Pretorius, Jakobus Maree
|
|
dc.contributor.author |
Huismans, H. (Henk), 1942-
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|
dc.contributor.author |
Theron, Jacques
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|
dc.date.accessioned |
2016-03-04T10:04:18Z |
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dc.date.issued |
2015-12 |
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dc.description.abstract |
Bluetongue virus (BTV), the type species of the genus Orbivirus within the family Reoviridae, has a genome consisting of 10 linear double-stranded RNA genome segments. Current reverse genetics approaches for engineering the BTV genome rely upon in vitro synthesis of capped RNA transcripts from cloned cDNA corresponding to viral genome segments. In an effort to expand the utility of BTV reverse genetics, we constructed a reverse genetics vector containing a T7 RNA polymerase promoter, hepatitis delta ribozyme sequence and T7 RNA polymerase terminator sequence. Viable virus was recovered following transfection of mammalian cells, expressing T7 RNA polymerase, with 10 plasmid constructs representing the cloned BTV-1 genome. Furthermore, the plasmid-based reverse genetics system was used successfully to isolate viable cross-serotype reassortant viruses and a mutant virus containing a defined mutation in the replicating viral genome. The new reverse genetics platform established here for BTV is likely applicable to other orbiviruses. |
en_ZA |
dc.description.embargo |
2016-12-30 |
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dc.description.librarian |
am2015 |
en_ZA |
dc.description.sponsorship |
A Grant (A0Y367) from the Tech-
nology Innovation Agency(TIA). |
en_ZA |
dc.description.uri |
http://www.elsevier.com/locate/yviro |
en_ZA |
dc.identifier.citation |
Pretorius, JM, Huismans, H & Theron, J 2015, 'Establishment of an entirely plasmid-based reverse genetics system for Bluetongue virus', Virology, vol. 486, pp. 71-77. |
en_ZA |
dc.identifier.issn |
0042-6822 (print) |
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dc.identifier.issn |
1096-0341 (online) |
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dc.identifier.other |
http://dx.doi.org/10.1016/j.virol.2015.09.004 |
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dc.identifier.uri |
http://hdl.handle.net/2263/51691 |
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dc.language.iso |
en |
en_ZA |
dc.publisher |
Elsevier |
en_ZA |
dc.rights |
© 2015 Elsevier Inc. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Virology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Virology, vol. 486, pp. 71-77. 2015. doi:10.1016/j.virol.2015.09.004. |
en_ZA |
dc.subject |
Orbivirus |
en_ZA |
dc.subject |
DsRNA |
en_ZA |
dc.subject |
Reverse genetics |
en_ZA |
dc.subject |
Reverse genetics plasmid |
en_ZA |
dc.subject |
T7 RNA Polymerase |
en_ZA |
dc.subject |
Genome modification |
en_ZA |
dc.subject |
Reassortment |
en_ZA |
dc.subject |
Bluetongue virus (BTV) |
en_ZA |
dc.title |
Establishment of an entirely plasmid-based reverse genetics system for Bluetongue virus |
en_ZA |
dc.type |
Postprint Article |
en_ZA |