Construction of Escherichia coli K‐12 in‐frame, single‐gene knockout mutants: the Keio collection
| Autor: | Takeshi Ara, Barry L. Wanner, Miki Baba, Tomoya Baba, Hirotada Mori, Miki Hasegawa, Yoshiko Okumura, Masaru Tomita, Kirill A. Datsenko, Yuki Takai |
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| Rok vydání: | 2006 |
| Předmět: |
E. coli/gene
Mutant Gene regulatory network Biology medicine.disease_cause Article General Biochemistry Genetics and Molecular Biology Escherichia coli knockout mutants medicine Coding region Gene bacterial functional genomics Genetics Internet Mutation Organisms Genetically Modified General Immunology and Microbiology Applied Mathematics systems biology Kanamycin essential gene Computational Theory and Mathematics Essential gene resources General Agricultural and Biological Sciences Gene Deletion Information Systems medicine.drug |
| Zdroj: | Molecular Systems Biology |
| ISSN: | 1744-4292 |
| Popis: | We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosomal genes and primers designed to create in-frame deletions upon excision of the resistance cassette. Of 4288 genes targeted, mutants were obtained for 3985. To alleviate problems encountered in high-throughput studies, two independent mutants were saved for every deleted gene. These mutants—the ‘Keio collection'—provide a new resource not only for systematic analyses of unknown gene functions and gene regulatory networks but also for genome-wide testing of mutational effects in a common strain background, E. coli K-12 BW25113. We were unable to disrupt 303 genes, including 37 of unknown function, which are candidates for essential genes. Distribution is being handled via GenoBase (http://ecoli.aist-nara.ac.jp/). |
| Databáze: | OpenAIRE |
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