C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/40 (2006.01) A61K 39/12 (2006.01) C07K 14/08 (2006.01) C12N 7/00 (2006.01) C12N 15/86 (2006.01) A61K 39/00 (2006.01)
Patent
CA 2264488
A system for the generation of live Birnavirus such as infectious bursal disease virus (IBDV), a segmented double-stranded (ds)RNA virus of the Birnavirdae family, using synthetic transcripts derived from cloned DNA has been developed. Independent full-length cDNA clones were constructed which contained the entire coding and non-coding regions of RNA segments A and B of IBDV, respectively. Synthetic RNAs of both segments were produced by in vitro transcription of linearized plasmids with T7 RNA polymerase. Transfection of Vero cells with combined plus-sense transcripts of both segments generated infectious virus as early as 36 hours post-transfection. The development of a reverse genetics system for dsRNA viruses will greatly facilitate studies of the regulation of viral gene expression pathogenesis, and design of a new generation of live and inactivated vaccines.
L'invention concerne un système servant à fabriquer un Birnavirus vivant, tel qu'un virus de bursite infectieuse (IBDV) - qui est un virus d'ARN bicaténaire segmenté de la famille Birnavirdae - à partir de transcrits synthétiques dérivés d'ADN cloné. Des clones d'ARNc entiers et indépendants sont fabriqués de façon à contenir les zones de codage et de non codage entières des segments d'ARN A et B de l'IBDV, respectivement. Les ARN synthétiques des deux segments ont été produits par transcription in vitro de plasmides linéarisés avec une polymérase d'ARN T7. La transfection de cellules Véro avec des transcrits combinés à sens positif des deux segments produit un virus infectieux seulement 36 heures après la transfection. Le développement d'un système génétique inverse de virus d'ARN bicaténaires facilitera grandement les recherches sur la régulation de la pathogenèse de l'expression de gènes viraux et la conception d'une nouvelle génération de vaccins vivants et rendus inactifs.
Mundt Egbert
Vakharia Vikram N.
Smart & Biggar
University Of Maryland - Biotechnology Institute
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