Method for identification of suitable fragmentation sites in...

Details Method for identification of suitable fragmentation sites in... Method for identification of suitable fragmentation sites in...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2542182

The invention concerns a combinatorial method for the generation of new split- protein sensors, and its application towards the (.beta./.alpha.)8-barrel enzyme N-(5.prime.-phosphoribosyl)-anthranilate isomerase Trp1p from Saccharomyces cerevisiae is demonstrated. The generated split-Trp protein sensors allow for the detection of protein-protein interactions in the cytosol as well as the membrane by enabling trp1 cells to grow on medium lacking tryptophan. This powerful selection thus complements the repertoire of the currently used split-protein sensors and provides a new tool for high- throughput interaction screening.

L'invention concerne une méthode de combinaison permettant de générer de nouveaux capteurs de protéines de division, et son utilisation avec l'enzyme N-(5.prime.-phosphoribosyl)-anthranilate isomérase Trp1p à réservoir (.beta./.alpha.)¿8? de Saccharomyces cerevisiae. Les capteurs des protéines Trp de division générés permettent de détecter des interactions protéine-protéine dans le cytosol, ainsi que la membrane, en permettant aux cellules trp1 de se développer sur un tryptophane sans milieu. Cette sélection vient ainsi complémenter le répertoire des capteurs de protéines de division actuellement utilisés, et permet d'obtenir un nouvel outil de criblage des interactions à haut rendement.

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