Method for the identification of essential genes and...

C - Chemistry – Metallurgy – 12 – Q

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Details

C12Q 1/68 (2006.01) C12N 15/10 (2006.01) C12Q 1/02 (2006.01) C12Q 1/18 (2006.01)

Patent

CA 2215870

The present invention relates to a method of identifying essential genes in a genome, based on an insertional mutagenesis of a population of cells or of DNA molecules and subjecting this population of cells or DNA molecules to an amplification process, whereby this total population of cells or DNA molecules which statistically represents at least one full insertionally mutated genome is amplified with at least two primer pairs and the extension products analysed, in order to distinguish essential genes from dispensable genes. The present invention is especially suited to the functional analysis of microbial genomes, and especially to haploid genomes.

L'invention concerne une méthode d'identification des gènes essentiels dans un génome, basée sur une mutagenèse insertionnelle d'une population de cellules ou de molécules d'ADN, cette dernière ou ces dernières étant ensuite soumises à un processus d'amplification. Cette population totale de cellules ou ces molécules d'ADN qui représentent statistiquement un génome ayant subi au moins une mutation insertionnelle complète sont amplifiées avec au moins deux paires d'amorce, et les produits d'extension sont analysés, afin de distinguer les gènes essentiels de ceux qui ne le sont pas. L'invention convient spécialement pour l'analyse fonctionnelle des génomes microbiens et, particulièrement, des génomes haploïdes.

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