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Transposon Insertion Site Sequencing in a Urinary Tract Model

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Proteus mirabilis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2021))

Abstract

Transposon sequencing (Tn-seq) is a technique that combines quantitative next-generation sequencing and a saturating transposon mutant library for an organism of interest, and ultimately allows for quantitation of the relative abundance of all of the mutants under a given condition, such as during experimental infection. The massively parallel sequencing capabilities of this technique provide a significant advance over more traditional methods of screening transposon mutant pools or individually determining the fitness contribution of genes of interest. Here, we describe a method for generating a genome-saturating transposon mutant library in Proteus mirabilis, determining the appropriate number of mutants for inoculation in an experimental infection model, preparing transposon insertion junctions for Illumina sequencing, and downstream analysis of mapped DNA sequencing reads for estimation of the contribution of each gene in the genome to fitness during infection.

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Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA

    Valerie S. Forsyth & Harry L. T. Mobley

  2. Department of Microbiology and Immunology, Jacobs School of Medicine and Biomedical Sciences, State University of New York at Buffalo, Buffalo, NY, USA

    Chelsie E. Armbruster

Authors
  1. Valerie S. Forsyth
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  2. Harry L. T. Mobley
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  3. Chelsie E. Armbruster
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Corresponding author

Correspondence to Chelsie E. Armbruster .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA

    Melanie M. Pearson

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Forsyth, V.S., Mobley, H.L.T., Armbruster, C.E. (2019). Transposon Insertion Site Sequencing in a Urinary Tract Model. In: Pearson, M. (eds) Proteus mirabilis. Methods in Molecular Biology, vol 2021. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9601-8_25

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  • DOI: https://doi.org/10.1007/978-1-4939-9601-8_25

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9600-1

  • Online ISBN: 978-1-4939-9601-8

  • eBook Packages: Springer Protocols

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