Genetics-assisted target evaluation in antibacterial drug...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/01 (2006.01) C12Q 1/04 (2006.01) C12Q 1/14 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2328497

The present invention provides a method, which is designated GATE (Genetics- Assisted Target Evaluation), for gene analysis in pathogenic bacteria. GATE functionally classifies genes according to their essential or nonessential character with respect to bacterial growth or pathogenicity. The technology uses a nonessential scorable gene as an internal standard against which the dispensability of a given target gene is assessed by gene disruption by Campbell-type integration. Functional analysis was performed on 26 genes in the Gram-positive pathogen Streptococcus pyogenes. Application of this method resulted in a clear identification of essential and nonessential loci: 16 of the 26 genes evaluated in S. pyogenes clearly encoded functions essential for bacterial growth, while the remaining 10 encoded functions that were nonessential. The successful application of GATE has also been demonstrated in Staphylococcus aureus.

L'invention concerne un procédé baptisé GATE (Genetics-Assisted Target Evaluation, ou "estimation de cibles assistée génétiquement"), destiné à l'analyse des gènes dans des bactéries pathogènes. Le procédé GATE permet de classer les gènes de manière fonctionnelle, selon leur caractère essentiel ou non essentiel en rapport avec la croissance bactérienne ou la pathogénie. La technique de l'invention s'appuie sur un gène non essentiel pouvant être soumis à la notation, utilisé en tant que standard interne par rapport auquel on évalue l'aptitude à la distribution d'un gène cible donné par disruption génique au moyen de l'intégration de type Campbell. Une analyse fonctionnelle a été effectuée sur 26 gènes du pathogène gram positif Streptococcus pyogenes. L'application de ce procédé a permis d'identifier clairement les loci essentiels et non essentiels: seize sur les vingt-six gènes évalués chez S. pyogenes codaient clairement les fonctions essentielles pour la croissance bactérienne, les dix autres fonctions codées n'étant pas essentielles. Le succès du procédé GATE a aussi été démontré vis-à-vis de Staphylococcus aureus.

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