Method for screening a proteasome activity modulating agent...

Details Method for screening a proteasome activity modulating agent... Method for screening a proteasome activity modulating agent...

G - Physics

01

N

G01N 33/68 (2006.01) C12N 1/16 (2006.01) C12N 15/81 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2626600

The invention relates to an in cellulo method for screening a proteasome activity modulating agent consisting a) in bringing a testable candidate agent into contact with recombinant yeast cells expressing in the core thereof a fusion protein comprising (i) a p21 polypeptide selected from p21WAF1/Cip1 and p21[6KR]WAF1/Cip1 polypeptides and (ii) at least one type of detectable protein, b) in quantifying said first detectable protein in the yeast cells at the end of at least one predetermined time interval after bringing the candidate agent into contact with the yeast cells, and c) in comparing the value obtained at the stage (b) with a control value obtained when the stage (a) is performed in the absence of the candidate agent.

L'invention a pour objet un procédé in cellulo pour le criblage d'agents modulant l'activité du protéasome, ledit procédé comprenant les étapes suivantes : a) mettre en contact un agent candidat à tester avec des cellules de levure recombinantes qui expriment dans leur noyau une protéine de fusion comprenant : (i) un polypeptide p21 choisi parmi le polypeptide p21WAF1/Cip1 et le polypeptide p21 [6KR]WAF1/Cip1 ;et (ii) au moins une protéine détectable. b) quantifier ladite première protéine détectable dans les cellules de levure, à la fin d'au moins une période de temps prédéterminée après la mise en contact de l'agent candidat avec lesdites cellules ; c) comparer la valeur obtenue à l'étape (b) avec une valeur témoin obtenue lorsque l'étape (a) est réalisée en l'absence de l'agent candidat.

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