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Gene Essentiality pp 45–65Cite as

Identification of Essential Genes and Synthetic Lethal Gene Combinations in Escherichia coli K-12

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1279))

Abstract

Here we describe the systematic identification of single genes and gene pairs, whose knockout causes lethality in Escherichia coli K-12. During construction of precise single-gene knockout library of E. coli K-12, we identified 328 essential gene candidates for growth in complex (LB) medium. Upon establishment of the Keio single-gene deletion library, we undertook the development of the ASKA single-gene deletion library carrying a different antibiotic resistance. In addition, we developed tools for identification of synthetic lethal gene combinations by systematic construction of double-gene knockout mutants. We introduce these methods herein.

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Acknowledgements

This work was supported by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and a grant from CREST, JST (Japan Science and Technology), NEDO (New Energy and Industrial Technology Development Organization), Tsuruoka City, Yamagata prefecture in Japan. B.L.W. is supported by the US National Science Foundation (grant number 106394).

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Authors and Affiliations

  1. Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0101, Japan

    Hirotada Mori, Katsushi Yokoyama, Rikiya Takeuchi, Wataru Nomura, Kazuichi Makishi, Yuta Otsuka & Hitomi Dose

  2. Transdiscplinary Research Integration Center, Mishima, Shizuoka, 411-8540, Japan

    Tomoya Baba

  3. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA, 02115, USA

    Barry L. Wanner

Authors
  1. Hirotada Mori
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  2. Tomoya Baba
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  3. Katsushi Yokoyama
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  4. Rikiya Takeuchi
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  5. Wataru Nomura
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  6. Kazuichi Makishi
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  7. Yuta Otsuka
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  8. Hitomi Dose
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  9. Barry L. Wanner
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Corresponding authors

Correspondence to Hirotada Mori or Barry L. Wanner .

Editor information

Editors and Affiliations

  1. Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, USA

    Long Jason Lu

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Mori, H. et al. (2015). Identification of Essential Genes and Synthetic Lethal Gene Combinations in Escherichia coli K-12. In: Lu, L. (eds) Gene Essentiality. Methods in Molecular Biology, vol 1279. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2398-4_4

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  • DOI: https://doi.org/10.1007/978-1-4939-2398-4_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2397-7

  • Online ISBN: 978-1-4939-2398-4

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