Towards the Development of a Minigenome Assay for Species A Rotaviruses.

Autor: Diebold O; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Zhou S; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Sharp CP; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Tesla B; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Chook HW; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Digard P; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK., Gaunt ER; Virology Division, Roslin Institute, University of Edinburgh, Easter Bush Campus, Midlothian EH25 9RG, UK.
Jazyk: angličtina
Zdroj: Viruses [Viruses] 2024 Aug 31; Vol. 16 (9). Date of Electronic Publication: 2024 Aug 31.
DOI: 10.3390/v16091396
Abstrakt: RNA virus polymerases carry out multiple functions necessary for successful genome replication and transcription. A key tool for molecular studies of viral RNA-dependent RNA polymerases (RdRps) is a 'minigenome' or 'minireplicon' assay, in which viral RdRps are reconstituted in cells in the absence of full virus infection. Typically, plasmids expressing the viral polymerase protein(s) and other co-factors are co-transfected, along with a plasmid expressing an RNA encoding a fluorescent or luminescent reporter gene flanked by viral untranslated regions containing cis -acting elements required for viral RdRp recognition. This reconstitutes the viral transcription/replication machinery and allows the viral RdRp activity to be measured as a correlate of the reporter protein signal. Here, we report on the development of a 'first-generation' plasmid-based minigenome assay for species A rotavirus using a firefly luciferase reporter gene.
Databáze: MEDLINE
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