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Transposon Mutagenesis in Streptococcus Species

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Microbial Transposon Mutagenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2016))

Abstract

Mutant libraries, generated by transposons and screened for various phenotypes, have led to many important discoveries regarding gene functions in various organisms. In this chapter we describe the use of plasmid pMN100, a transposon vector constructed to perform in vivo transposition primarily in oral streptococci. Compared to in vitro transposition systems the conditional replicative features of the plasmid, and the inducible expression of the mariner Himar1 transposase, makes pMN100 particularly useful for bacterial strains showing a low transformation frequency. We outline how to transform plasmid pMN100 into Streptococcus mutans, carry out transposon mutagenesis, and determine the chromosomal location of inserted transposons. It is our prospect that the protocols can be used as guidelines for transposon mutagenesis in S. mutans as well as other species of streptococci.

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Acknowledgments

This work was supported by grants from the Danish Council for Independent Research and the Lundbeck Foundation.

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Authors and Affiliations

  1. Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, Costerton Biofilm Center, University of Copenhagen, Copenhagen, Denmark

    Martin Nilsson, Michael Givskov & Tim Tolker-Nielsen

  2. Singapore Center for Environmental Life Sciences Engineering, Nanyang Technological University, Singapore, Singapore

    Michael Givskov

Authors
  1. Martin Nilsson
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  2. Michael Givskov
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  3. Tim Tolker-Nielsen
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Corresponding author

Correspondence to Tim Tolker-Nielsen .

Editor information

Editors and Affiliations

  1. Department of Food Science, Center for Food Safety, University of Arkansas, Fayetteville, AR, USA

    Steven C. Ricke

  2. Department of Food Science and Technology, Oregon State University, Corvallis, OR, USA

    Si Hong Park

  3. Department of Food Science, University of Arkansas, Fayetteville, AR, USA

    Morgan L. Davis

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Nilsson, M., Givskov, M., Tolker-Nielsen, T. (2019). Transposon Mutagenesis in Streptococcus Species. In: Ricke, S., Park, S., Davis, M. (eds) Microbial Transposon Mutagenesis. Methods in Molecular Biology, vol 2016. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9570-7_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9570-7_4

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9569-1

  • Online ISBN: 978-1-4939-9570-7

  • eBook Packages: Springer Protocols

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