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Genetic Transformation and Complementation

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Borrelia burgdorferi

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1690))

Abstract

The disciplines of Borrelia (Borreliella) burgdorferi microbiology and Lyme disease pathogenesis have come to depend on the genetic manipulation of the spirochete. Generating mutants in these recalcitrant bacteria, while not straightforward, is routinely accomplished in numerous laboratories, although there are several crucial caveats to consider. This chapter describes the design of basic molecular genetic experiments as well as the detailed methodologies to prepare and transform competent cells, select for and isolate transformants, and complement or genetically restore mutants.

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Acknowledgments

We thank Phil Stewart for thoughtful review of the manuscript, Rich Marconi, Patti Rosa, Tom Schwan, and Kit Tilly for advice during development of the original protocol in the early 1990s, Frank Yang for suggesting the cloning-by-limiting-dilution protocol, and Christian Eggers, Mike Gilbert, Meghan Lybecker and the other members, past and present, of our laboratory for useful discussions. Genetic transformation and complementation experiments in our laboratory are supported by National Institutes of Health grant AI051486 (to D.S.S.).

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Authors and Affiliations

  1. Division of Biological Sciences, University of Montana, Missoula, MT, USA

    D. Scott Samuels, Dan Drecktrah & Laura S. Hall

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  1. D. Scott Samuels
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  2. Dan Drecktrah
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Correspondence to D. Scott Samuels .

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Editors and Affiliations

  1. Department of Veterinary Medicine, University of Maryland, College Park, Maryland, USA

    Utpal Pal

  2. Department of Microbiology, Ondokuz Mayis University, Samsun, Turkey

    Ozlem Buyuktanir

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Samuels, D.S., Drecktrah, D., Hall, L.S. (2018). Genetic Transformation and Complementation. In: Pal, U., Buyuktanir, O. (eds) Borrelia burgdorferi. Methods in Molecular Biology, vol 1690. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7383-5_15

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  • DOI: https://doi.org/10.1007/978-1-4939-7383-5_15

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