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The Bacterial Nucleoid pp 39–49Cite as

Transposon Insertion Site Sequencing for Synthetic Lethal Screening

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1624))

Abstract

Transposon insertion site sequencing (TIS) permits genome-wide, quantitative fitness assessment of individual genomic loci. In addition to the identification of essential genes in given growth conditions, TIS enables the elucidation of genetic networks such as synthetic lethal or suppressor gene combinations. Therefore, TIS becomes an exceptionally powerful tool for the high-throughput determination of genotype-phenotype relationships in bacteria. Here, we describe a protocol for the generation of high-density transposon insertion libraries and subsequent preparation of DNA samples for Illumina sequencing using the Gram-negative bacterium Vibrio cholerae as an example.

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Authors and Affiliations

  1. Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Université Paris Sud, 1 Avenue de la Terrasse, 91198, Gif sur Yvette, France

    Yoshiharu Yamaichi

  2. Department of Microbiology, Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY, 14853, USA

    Tobias Dörr

Authors
  1. Yoshiharu Yamaichi
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  2. Tobias Dörr
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Corresponding authors

Correspondence to Yoshiharu Yamaichi or Tobias Dörr .

Editor information

Editors and Affiliations

  1. UMR CNRS 7241, INSERM U1050, CIRB, Collège de France, Paris, France

    Olivier Espéli

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Yamaichi, Y., Dörr, T. (2017). Transposon Insertion Site Sequencing for Synthetic Lethal Screening. In: Espéli, O. (eds) The Bacterial Nucleoid. Methods in Molecular Biology, vol 1624. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7098-8_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7098-8_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7097-1

  • Online ISBN: 978-1-4939-7098-8

  • eBook Packages: Springer Protocols

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