Cited by 7,858Bacterial Genetics and BiotechnologyMicrobial Metabolic Engineering and BioproductionCRISPR and Genetic Engineering

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Construction of Escherichia coli K‐12 in‐frame, single‐gene knockout mutants: the Keio collection

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Baba, Tomoya, Ara, Takeshi, Hasegawa, Miki, Takai, Yuki, Okumura, Yoshiko, Baba, Miki, Datsenko, Kirill A, Tomita, Masaru, Wanner, Barry L, Mori, Hirotada·2006·Molecular Systems Biology

DOI 10.1038/msb4100050PMC PMC1681482

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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/).

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Authors: Baba, T., Ara, T., Hasegawa, M., Takai, Y., Okumura, Y., Baba, M., Datsenko, K. A., Tomita, M., Wanner, B. L., & Mori, H.
Affiliations: Institute for Advanced Biosciences, Keio University (JP)
Japan Science and Technology Agency (JP)
Purdue University West Lafayette (US)
Corresponding author: Wanner, Barry L
Mori, Hirotada
ORCID: Baba, Tomoya
Ara, Takeshi
Hasegawa, Miki
Takai, Yuki
Tomita, Masaru
Wanner, Barry L
Mori, Hirotada
Venue: Molecular Systems Biology
Publisher: Springer Science and Business Media LLC
Published: 2006-02-21
Volume: 2
Issue: 1
Pages: 2006.0008-2006.0008
ISSN: 1744-4292 (ISSN-L)
DOI: 10.1038/msb4100050
PMC: PMC1681482
OpenAlex: W2111967267
Cited by: 7,858
References: 196
Citations by year: 2026: 331 · 2025: 452 · 2024: 489 · 2023: 429 · 2022: 458 · 2021: 490 · 2020: 502 · 2019: 451 · 2018: 454 · 2017: 464 · 2016: 394 · 2015: 435 · 2014: 451 · 2013: 415 · 2012: 376
FWCI: 117.12
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: www.springer.com/tdm (tdm)
www.springer.com/tdm (vor)
Locations: Molecular Systems Biology (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: Keio University
National Institutes of Health
National Institute of General Medical Sciences (GM 62662, R01GM062662)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Keywords: Keio collection, Escherichia coli K-12, single gene deletion, non-essential genes, kanamycin resistance cassette, gene inactivation, gene function analysis, gene regulatory networks, essential genes
MeSH terms: Escherichia coli, Mutation, Gene Deletion, Internet, Organisms, Genetically Modified

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Construction of Escherichia coli K‐12 in‐frame, single‐gene knockout mutants: the Keio collection

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