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Quantitative Genetic Screens for Mapping Bacterial Pathways and Functional Networks

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Mapping Genetic Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2381))

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Abstract

Escherichia coli synthetic genetic array (eSGA) screening procedure enables high-throughput systematic mapping of pairwise genetic interactions in E. coli. The eSGA method exploits E. coli’s rapid growth, its ease of genetic manipulation, and efficient genetic exchange via conjugation. Replica pinning is used to grow and mate arrayed sets of single gene mutant strains as well as to select double mutants en masse. Strain fitness, which is the eSGA readout, is determined by the digital imaging of the plates and subsequent colony size measurements. Comparing single and double mutant colony sizes then allows for identifying interacting genes. Using eSGA on a global or a smaller process-centric scale can help reveal gene functions and reconstruct genetic interaction networks with known and novel connections between genes and pathways.

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Acknowledgments

AG is a recipient of Canadian Institute of Health Research Postdoctoral Fellowship. This work was supported by a grant from the National Sciences and Engineering Research Council of Canada to M.B (DG-20234).

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Authors and Affiliations

  1. Department of Biochemistry, Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK, Canada

    Alla Gagarinova

  2. Department of Biochemistry, University of Regina, Regina, SK, Canada

    Ali Hosseinnia & Mohan Babu

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  1. Alla Gagarinova
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  2. Ali Hosseinnia
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  3. Mohan Babu
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Correspondence to Alla Gagarinova or Mohan Babu .

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  1. University of Saskatchewan, Saskatoon, SK, Canada

    Franco Joseph Vizeacoumar

  2. University of Saskatchewan, Saskatoon, SK, Canada

    Andrew Freywald

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Gagarinova, A., Hosseinnia, A., Babu, M. (2021). Quantitative Genetic Screens for Mapping Bacterial Pathways and Functional Networks. In: Vizeacoumar, F.J., Freywald, A. (eds) Mapping Genetic Interactions. Methods in Molecular Biology, vol 2381. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1740-3_1

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  • DOI: https://doi.org/10.1007/978-1-0716-1740-3_1

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