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Bacteriophage Isolation and Characterization: Phages of Escherichia coli

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Horizontal Gene Transfer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2075))

Abstract

Here we introduce methods for the detection, enumeration, and isolation of bacteriophages from Escherichia coli. In bacteria, horizontal gene transfer may be mediated by virulent and temperate phages. Strict virulent phages, able to propagate in a suitable strain following the lytic pathway, can be isolated directly from different natural environments. In temperate phages, the lytic cycle must be activated, and phages are detected after their induction. In both cases, detection is based on the production of visible plaques in a confluent lawn of the host strain using a double agar layer method. Further purification and characterization are achieved by density gradients, electron microscopy studies, and genomic analysis. This straightforward methodology can be applied to the detection, enumeration, and isolation of bacteriophages from any bacterial species, using the appropriate host strain, media, and culture conditions.

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Authors and Affiliations

  1. Department of Microbiology, University of Barcelona, Barcelona, Spain

    Juan Jofre & Maite Muniesa

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  1. Juan Jofre
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  2. Maite Muniesa
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Correspondence to Maite Muniesa .

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Editors and Affiliations

  1. Instituto de Biomedicina y Biotecnología de Cantabria, Universidad de Cantabria-CSIC, Santander, Cantabria, Spain

    Fernando de la Cruz

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Jofre, J., Muniesa, M. (2020). Bacteriophage Isolation and Characterization: Phages of Escherichia coli. In: de la Cruz, F. (eds) Horizontal Gene Transfer. Methods in Molecular Biology, vol 2075. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9877-7_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9877-7_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9876-0

  • Online ISBN: 978-1-4939-9877-7

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