Target assisted iterative screening (tais) : a novel...

G - Physics – 01 – N

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G01N 33/53 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2462732

This invention provides a new in vitro screening method for the detection of protein-protein and other interactions. The method has been developed and applied to a commercial cDNA library to search for novel protein-protein interactions. PDZ, WW and SH3 domains from PSD95, Nedd4, Src, Abl and Crk proteins were used as test targets. 12 novel putative and 2 previously reported interactions were identified for 6 protein interaction modules in test screens. The novel screening format, dubbed TAIS (target-assisted iterative screening), provides an alternative platform to existing technologies for a pair-wise characterization of protein-protein, and other, interactions.

L'invention concerne un nouveau procédé de criblage in vitro servant à déceler des interactions protéine-protéine et autres. Ledit procédé a été mis au point et appliqué à une banque d'ADNc commerciale pour rechercher de nouvelles interactions protéine-protéine. Les domaines PDZ, WW et SH3 des protéines PSD95, Nedd4, Src, Abl et Crk ont été utilisés comme cibles d'essai. Dans les criblages tests, 12 interactions estimées nouvelles et 2 interactions précédemment relevées ont été identifiées pour 6 modules d'interactions de protéines. Le nouveau format de criblage, le TAIS (criblage itératif assisté par cibles) doublé, constitue une plate-forme alternative aux techniques existantes pour la caractérisation par paires d'interactions protéine-protéine ou d'autres interactions.

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