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Random Mutagenesis Strategies for Campylobacter and Helicobacter Species

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In Vitro Mutagenesis Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 634))

Abstract

Campylobacterand Helicobacterspecies are important pathogens in man and animals. The study of their virulence and physiology has been difficult due to the lack of tractable genetic tools, since many of the techniques established in Escherichia coliand related species were found to be non-functional in Campylobacterand Helicobacterspecies. The advent of functional genomics techniques in the last decade has been accompanied by the development of genetic tools, which take advantage of specific features of Campylobacterand Helicobacter, like natural transformation. This has allowed for the construction of random mutant libraries based on in vitro transposition or ligated loops followed by natural transformation and recombination, thus circumventing selection against sequences when cloning or passaging libraries through E. coli. Uses of the techniques have been in the study of motility, gene expression, and gene essentiality. In this chapter, we discuss the approaches and techniques used for the construction of random mutant libraries in both Campylobacterand Helicobacter.

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Acknowledgements

The Campylobacterresearch in the Institute of Food Research is supported by the Core Strategic Grant from the Biotechnology and Biological Sciences Research Council (BBSRC). We thank Dr Jerry Wells, Dr Kay Fowler, and other past and present members of our laboratories for their contributions to the development of these protocols.

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Authors and Affiliations

  1. Institute of Food Research, Norwich, UK

    Duncan J. H. Gaskin & Arnoud H. M. van Vliet

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  1. Duncan J. H. Gaskin
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  2. Arnoud H. M. van Vliet
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  1. Stratagene, North Torrey Pines Rd. 11011, La Jolla, 92037, USA

    Jeff Braman

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Gaskin, D.J.H., van Vliet, A.H.M. (2010). Random Mutagenesis Strategies for Campylobacter and Helicobacter Species. In: Braman, J. (eds) In Vitro Mutagenesis Protocols. Methods in Molecular Biology, vol 634. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-652-8_3

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  • DOI: https://doi.org/10.1007/978-1-60761-652-8_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-651-1

  • Online ISBN: 978-1-60761-652-8

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