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Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis

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Microbial Transposon Mutagenesis

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

Abstract

While transposon mutagenesis has been developed for Mycobacterium avium subspecies paratuberculosis (Map), relatively few laboratories have adopted this important genetic tool to examine gene function and essentiality. Here we describe the construction of a Map transposon library using the Himar1 mariner transposon, but concepts can also be applied to the Tn5367 transposon, which has also been used by our group. Delivery of the transposon is by a temperature-sensitive phagemid, ϕMycoMarT7, and plating transductants requires patience and specialized media due to length of incubation required to observe colonies. Several transposon mutants obtained from these libraries have been tested in vaccine and pathogenesis studies. By providing the following detailed protocol herein, we expect to demystify the procedure and encourage additional investigators to incorporate transposon mutagenesis in their studies on Johne’s disease.

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Acknowledgments

This work was supported by the USDA-Agricultural Research Service and the USDA-National Institute of Food and Agriculture grant award 2013-67015.

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Authors and Affiliations

  1. USDA-ARS-National Animal Disease Center, Ames, IA, USA

    John P. Bannantine

  2. School of Veterinary Medicine and Biomedical Sciences, University of Nebraska, Lincoln, NE, USA

    Denise K. Zinniel & Raúl G. Barletta

Authors
  1. John P. Bannantine
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  2. Denise K. Zinniel
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  3. Raúl G. Barletta
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Corresponding author

Correspondence to John P. Bannantine .

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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Bannantine, J.P., Zinniel, D.K., Barletta, R.G. (2019). Transposon Mutagenesis in Mycobacterium avium Subspecies Paratuberculosis . 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_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9570-7_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9569-1

  • Online ISBN: 978-1-4939-9570-7

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