Abstract

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/).

Keywords

BiologyMutantEscherichia coliGene knockoutGeneticsFrame (networking)GeneComputational biologyComputer science

MeSH Terms

Escherichia coliGene DeletionInternetMutationOrganismsGenetically Modified

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Publication Info

Year
2006
Type
article
Volume
2
Issue
1
Pages
2006.0008-2006.0008
Citations
7987
Access
Closed

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Cite This

Tomoya Baba, Takeshi Ara, Miki Hasegawa et al. (2006). Construction of <i>Escherichia coli</i> K‐12 in‐frame, single‐gene knockout mutants: the Keio collection. Molecular Systems Biology , 2 (1) , 2006.0008-2006.0008. https://doi.org/10.1038/msb4100050

Identifiers

DOI
10.1038/msb4100050
PMID
16738554
PMCID
PMC1681482

Data Quality

Data completeness: 90%