Screening system for antibiotics

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/18 (2006.01) C12N 15/00 (2006.01)

Patent

CA 2408903

The invention provides a method of detecting in a sample the activity of an antibiotic which affects cell integrity, which method comprises the steps of: a) providing a transformed microorganism which comprises a nucleic acid encoding a promoter operably linked to a heterologous reporter gene capable of causing a detectable signal; b) contacting the sample with the transformed microorganism; c) observing said microorganism for said detectable signal, wherein the promoter is regulated by a two-component signal transduction system, which signal components are: i) a receptor responsive to changes in the cell envelope or membrane of the microorganism, ii) a trans-acting factor which is activated in response to stimulation by the receptor, and which regulates the promoter. Preferred signal components are derived from CseB and CseC which regulate a sigE promoter. Also provided are corresponding microorganisms, systems, kits, and processes of use of these things.

La présente invention concerne une méthode de détection, dans un échantillon, de l'activité d'un antibiotique affectant l'intégrité cellulaire. Cette méthode consiste à: a) fournir un micro-organisme transformé qui contient un acide nucléique codant un promoteur liée de manière fonctionnelle à un gène rapporteur hétérologue pouvant émettre un signal détectable; b) mettre en contact l'échantillon avec le micro-organisme transformé; c) observer ledit micro-organisme pour détecter ledit signal détectable, le promoteur étant régulé par un système de transduction de signal à deux composantes, les composantes du signal étant: 1) un récepteur sensible aux modifications de la membrane cellulaire du micro-organisme, 2) un facteur trans-activateur qui s'active en réponse à la stimulation du récepteur, et qui régule le promoteur. Les composantes du signal préférées sont obtenues à partir de CseB et CseC qui régulent un promoteur sigE. Par ailleurs, cette invention concerne des micro-organismes, des systèmes, et des kits correspondants ainsi que leurs procédés d'utilisation.

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