Genetic selection system to identify proteases, protease...

G - Physics – 01 – N

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G01N 33/53 (2006.01)

Patent

CA 2623694

The present invention concerns a tester protein for identifying and/or monitoring protease activity in a cellular assay suitable for high throughput screenings by growth selection, wherein the tester polypeptide is a non- regulatory protein carrying a protease cleavage sequence. Upon co-expression of the protease recognizing said cleavage sequence the tester protein is inactivated, which influences the growth and/or survival of the host cells under the chosen conditions. However, in the presence of protease inhibitor the growth phenotype is reversed. The system can be used to identify proteases, protease inhibitors, and protease cleavage sites.

La présente invention a trait à une protéine d'essai pour l'identification et/ou le suivi de l'activité de protéase dans un dosage cellulaire appropriée pour des criblages à haut rendement par la sélection de croissance, dans lesquels un polypeptide d'essai est une protéine non régulatrice portant une séquence de clivage de la protéase. Lors de la co-expression de la protéase reconnaissant ladite séquence de clivage, la protéine d'essai est inactivée, ce qui influence la croissance et/ou la survie des cellules hôtes dans les conditions prédéterminées. Cependant, en présence d'inhibiteur de la protéase le phénotype de croissance est inversé. Le système peut être utilisé pour l'identification de protéases, d'inhibiteurs de protéases, et des sites de clivage de protéases.

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