C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/10 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 2472072
The invention concerns an exhaustive method for detecting pathogenic bacteria genes, in particular Nm genes, expressing a desired phenotype, characterized in that it consists in: using a bank of mutants generated from a given bacterial strain so that at least 70 % of the non-essential genes, and in particular at least 80 %, even more than 90 %, are mutagenized by inserting a transposon in a reading phase; then contacting the mutants, either individually, or in groups, with an environment, such as a medium, an animal or cells, capable of interacting with the mutant bacteria expressing the desired phenotype; recovering, when groups are used, the bacteria which have not reacted with the desired phenotype; identifying the mutated genes of said bacteria and verifying whether they are involved in said phenotype. The invention is useful, in particular, as anti-pathogenicity targets, which consists in inhibiting Neisseria meningitidis growth in vivo in the serum, for developing antibiotics, for screening and manufacturing medicines designed to open the blood brain barrier to active principles, and for preparing vaccines.
L'invention vise un procédé de détection exhaustif de gènes de bactéries pathogènes, en paticulier deNm, exprimant un phénotype recherché, caractérisé en ce que :- on utilise une banque de mutants générée à partir d'une souche bactérienne donnée de manière à ce qu'au moins 70% des gènes non essentiels, et notamment au moins 80%, voire plus de 90%, soient mutagénisés par insertion d'un transposon dans une phase de lecture, - on met ensuite en contact les mutants, soit individuellement, soit en pools, avec un environnement, tel que milieu, animal ou cellules, capable d'interagir avec les bactéries mutantes exprimant le phénotype recherché,- on récupère, dans le cas de l'utilisation de pools, les bactéries n'ayant pas interagei avec le phénotype recherché, - on identifie les gènes mutés de ces bactéries et on vérifie leur implication dans ledit phénotype. Applications, notamment, comme cibles d'antipathogénicité, consistant à inhiber la croissance de Neisseria meningitidis in vivo dans le sérum, pour développer des antibiotiques, pour cribler et fabriquer des médicaments permettant l'ouverture de la barrière hématoencéphalique à des principes actifs, et élaborer des vaccins.
Nassif Xavier
Pelicic Vladimir
Institut National de La Sante Et de La Recherche Medicale (i.n.s
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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