Identification of genes which allow a microorganism to adapt...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/31 (2006.01) A61K 31/70 (2006.01) A61K 35/74 (2006.01) A61K 39/00 (2006.01) A61K 39/112 (2006.01) C07K 14/21 (2006.01) C07K 14/255 (2006.01) C07K 14/315 (2006.01) C07K 14/38 (2006.01) C12N 9/52 (2006.01) C12N 15/10 (2006.01) C12N 15/57 (2006.01) C12Q 1/04 (2006.01) C12Q 1/18 (2006.01) C12Q 1/68 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2206515

A method for identifying a microorganism having a reduced adaptation to a particular environment comprising the steps of: (1) providing a plurality of microorganisms each of which is independently mutated by the insertional inactivation of a gene with a nucleic acid comprising a unique marker sequence so that each mutant contains a different marker sequence, or clones of the said microorganism; (2) providing individually a stored sample of each mutant produced by step (1) and providing individually stored nucleic acid comprising the unique marker sequence from each individual mutant; (3) introducing a plurality of mutants produced by step (1) into the said particular environment and allowing those microorganisms which are able to do so to grow in the said environment; (4) retrieving microorganisms from the said environment or a selected part thereof and isolating the nucleic acid from the retrieved microorganisms; (5) comparing any marker sequences in the nucleic acid isolated in step (4) to the unique marker sequence of each individual mutant stored as in step (2); and (6) selecting an individual mutant which does not contain any of the marker sequences as isolated in step (4).

Procédé d'identification d'un micro-organisme dont l'adaptation à un environnement particulier est réduite, consistant à: (1) obtenir une pluralité de micro-organismes dont chacun est soumis à une mutation indépendante par l'inactivation insertionnelle d'un gène au moyen d'un acide nucléique comprenant une séquence de marquage unique, de sorte que chaque mutant contienne une séquence de marquage différente ou des clones dudit micro-organisme; (2) prendre un échantillon individuel préalablement stocké de chaque mutant obtenu à l'étape (1) et extraire un acide nucléique stocké comprenant la séquence de marquage unique de chaque mutant individuel; (3) introduire une pluralité de mutants obtenus à l'étape (1) dans ledit environnement particulier et laisser les micro-organismes qui en sont capables s'y développer; (4) récupérer les micro-organismes dudit environnement ou d'une partie sélectionnée de ce dernier et isoler l'acide nucléique des micro-organismes récupérés; (5) comparer les séquences de marquage dans l'acide nucléique isolé à l'étape (4) à la séquence de marquage unique de chaque mutant individuel stocké à l'étape (2); et (6) sélectionner un mutant individuel qui ne contienne aucune des séquences de marquage isolées à l'étape (4).

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