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Replication and packaging of an infectious bursal disease virus segment A-derived minigenome.

Abstract : A synthetic cytomegalovirus (CMV) promoter-driven cDNA minigenome containing the enhanced green fluorescence protein (EGFP) gene as a reporter was derived from the genomic segment A of infectious bursal disease virus (IBDV). The 5'-end of the minigenome was fused to the transcription start site of the immediate early CMV promoter, and the hepatitis delta virus (HDV) ribozyme sequence was added at its 3'-end. We show that co-transfection of the minigenome with a plasmid encoding the IBDV RNA-dependent RNA polymerase VP1, results in a consistent increase of the EGFP expression, as measured by fluorescence microscopy and flow cytometry assays. Replication of the minigenome-derived transcript was evidenced by real-time RT-PCR analyses targeted to both the plus- and minus-sense strands. When cells were infected with IBDV and transfected with the plasmid carrying the minigenome, the minigenome was packaged and EGFP was found to be expressed in a second cycle of infection. These results show the potential use of this system as a new tool to characterize IBDV replication and genome packaging.
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Submitted on : Friday, July 1, 2011 - 12:43:56 PM
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N. Ben Abdeljelil, B. Delmas, H. Mardassi. Replication and packaging of an infectious bursal disease virus segment A-derived minigenome.. Virus Research, Elsevier, 2008, 136 (1-2), pp.146-51. ⟨10.1016/j.virusres.2008.05.002⟩. ⟨pasteur-00605376⟩



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