| Autor: |
Alexander AJT; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Confort MP; IVPC UMR754, INRAE, Univ Lyon, Université Claude Bernard Lyon1, EPHE, PSL Research University, F-69007 Lyon, France., Desloire S; IVPC UMR754, INRAE, Univ Lyon, Université Claude Bernard Lyon1, EPHE, PSL Research University, F-69007 Lyon, France., Dunlop JI; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Kuchi S; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Sreenu VB; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Mair D; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Wilkie GS; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Da Silva Filipe A; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Brennan B; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK., Ratinier M; IVPC UMR754, INRAE, Univ Lyon, Université Claude Bernard Lyon1, EPHE, PSL Research University, F-69007 Lyon, France., Arnaud F; IVPC UMR754, INRAE, Univ Lyon, Université Claude Bernard Lyon1, EPHE, PSL Research University, F-69007 Lyon, France., Kohl A; MRC-University of Glasgow Centre for Virus Research, Glasgow G61 1QH, Scotland, UK. |
| Abstrakt: |
Toscana virus (TOSV) is a Phlebovirus in the Phenuiviridae family, order Bunyavirales , found in the countries surrounding the Mediterranean. TOSV is an important cause of seasonal acute meningitis and encephalitis within its range. Here, we determined the full sequence of the TOSV strain 1500590, a lineage A virus obtained from an infected patient (Marseille, 2007) and used this in combination with other sequence information to construct functional cDNA plasmids encoding the viral L, M, and S antigenomic sequences under the control of the T7 RNA promoter to recover recombinant viruses. Importantly, resequencing identified two single nucleotide changes to a TOSV reference genome, which, when corrected, restored functionality to the polymerase L and made it possible to recover infectious recombinant TOSV (rTOSV) from cDNA, as well as establish a minigenome system. Using reverse genetics, we produced an NSs-deletant rTOSV and also obtained viruses expressing reporter genes instead of NSs. The availability of such a system assists investigating questions that require genetic manipulation of the viral genome, such as investigations into replication and tropism, and beyond these fundamental aspects, also the development of novel vaccine design strategies. |
