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Using Genome Scale Mutant Libraries to Identify Essential Genes

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Essential Genes and Genomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2377))

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Abstract

Identification of essential genes is key to understanding the required processes and gene products of organisms under one or more conditions. Transposon sequencing (Tn-seq) has been used to predict essential genes or ones that conditionally impact fitness in a wide variety of organisms. Here, we describe the generation of genome-scale mutant libraries and the analysis of Tn-seq data to identify essential genes from cultures grown in a single condition as well as those that are conditionally important by analyzing the behavior of these mutant libraries in different growth environments. While we illustrate the approach using data derived from Tn-seq analysis of the α-proteobacteria Rhodobacter sphaeroides and Zymomonas mobilis, the protocols and systems we describe should be generally applicable to a variety of organisms.

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Acknowledgements

This material is based upon work supported by the Great Lakes Bioenergy Research Center, U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research under Award Number DE-SC0018409.

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Authors and Affiliations

  1. Wisconsin Energy Institute and Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI, USA

    Kevin S. Myers, Piyush Behari Lal, Daniel R. Noguera & Timothy J. Donohue

  2. Department of Civil & Environmental Engineering, University of Wisconsin-Madison, Madison, WI, USA

    Daniel R. Noguera

  3. Department of Bacteriology, University of Wisconsin-Madison, Madison, WI, USA

    Timothy J. Donohue

Authors
  1. Kevin S. Myers
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  2. Piyush Behari Lal
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  3. Daniel R. Noguera
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  4. Timothy J. Donohue
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Corresponding author

Correspondence to Timothy J. Donohue .

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Editors and Affiliations

  1. Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI, USA

    Ren Zhang

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Myers, K.S., Behari Lal, P., Noguera, D.R., Donohue, T.J. (2022). Using Genome Scale Mutant Libraries to Identify Essential Genes. In: Zhang, R. (eds) Essential Genes and Genomes. Methods in Molecular Biology, vol 2377. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1720-5_12

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  • DOI: https://doi.org/10.1007/978-1-0716-1720-5_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1719-9

  • Online ISBN: 978-1-0716-1720-5

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