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 7/04 (2006.01) C12N 15/86 (2006.01) A61K 39/00 (2006.01)
Patent
CA 2324478
A system for the generation of live, nonpathogenic infectious pancreatic necrosis virus (IPNV), 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 coding and non-coding regions of RNA segments A and B of IPNV, respectively. Segment A was modified to prevent the expression of NS protein. Synthetic RNAs of both segments were produced by in vitro transcription of linearized plasmids with T7 RNA polymerase. Transfection of CHSE cells with combined plus-sense transcripts of both segments generated infectious virus. The development of a system for producing NS protein deficient IPNV will greatly facilitate studies of viral pathogenesis, and the development of live attenuated vaccines for IPNV.
On a mis au point un système permettant de générer un virus de nécrose pancréatique infectieux, non pathogène, vivant (IPNV), un virus d'ARN/ds segmenté, double-brin qui utilisent des produits de transcription synthétiques, dérivés de l'ADN clonée. On a élaboré des clones d'ADNc indépendants, pleine longueur, contenant les régions codante et non codante des segments d'ARN A et B respectivement, de l'IPNV. Un segment A a été modifié pour empêcher l'expression de la protéine NS. Les ARN synthétiques des deux segments ont été produits par transcription in vitro de plasmides linéarisés avec l'ARN-polymérase T7. La transfection de cellules CHSE avec des produits de transcription combinés sens positif des deux segments a généré un virus infectieux. La mise au point d'un système permettant de produire un IPNV privé de protéine NS facilite considérablement les études sur la pathogénèse virale et l'élaboration de vaccins atténués vivants pour IPNV.
Vakharia Vikram N.
Yao Kun
Blake Cassels & Graydon Llp
University Of Maryland College Park
University Of Maryland - Biotechnology Institute
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