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Clostridium difficile pp 143–165Cite as

Characterization of Functional Prophages in Clostridium difficile

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1476))

Abstract

Bacteriophages (phages) are present in almost, if not all ecosystems. Some of these bacterial viruses are present as latent “prophages,” either integrated within the chromosome of their host, or as episomal DNAs. Since prophages are ubiquitous throughout the bacterial world, there has been a sustained interest in trying to understand their contribution to the biology of their host. Clostridium difficile is no exception to that rule and with the recent release of hundreds of bacterial genome sequences, there has been a growing interest in trying to identify and classify these prophages. Besides their identification in bacterial genomes, there is also growing interest in determining the functionality of C. difficile prophages, i.e., their capacity to escape their host and reinfect a different strain, thereby promoting genomic evolution and horizontal transfer of genes through transduction, for example of antibiotic resistance genes. There is also some interest in using therapeutic phages to fight C. difficile infections.

The objective of this chapter is to share with the broader C. difficile research community the expertise we developed in the study of C. difficile temperate phages. In this chapter, we describe a general “pipeline” comprising a series of experiments that we use in our lab to identify, induce, isolate, propagate, and characterize prophages. Our aim is to provide readers with the necessary basic tools to start studying C. difficile phages.

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Authors and Affiliations

  1. Département de Microbiologie et d’infectiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, 3201 rue Jean Mignault, Sherbrooke, QC, Canada, J1E 4K8

    Ognjen Sekulović & Louis-Charles Fortier

Authors
  1. Ognjen Sekulović
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  2. Louis-Charles Fortier
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Correspondence to Louis-Charles Fortier .

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Editors and Affiliations

  1. Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, United Kingdom

    Adam P. Roberts

  2. Department of Microbial Diseases, UCL Eastman Dental Institute, University College London, London, United Kingdom

    Peter Mullany

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Sekulović, O., Fortier, LC. (2016). Characterization of Functional Prophages in Clostridium difficile . In: Roberts, A., Mullany, P. (eds) Clostridium difficile. Methods in Molecular Biology, vol 1476. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6361-4_11

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  • DOI: https://doi.org/10.1007/978-1-4939-6361-4_11

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