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Creation of Staphylococcal Mutant Libraries Using Transposon Tn917

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The Genetic Manipulation of Staphylococci

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1373))

Abstract

Non-directed mutagenesis of the staphylococcal genome is a global approach that can be used to identify the genetic basis for phenotypes of interest as well as for identifying regulators of gene expression. One such approach that has been widely used in the study of S. aureus and S. epidermidis is transposon Tn917 mutagenesis to generate random libraries of mutants. This chapter describes the use of plasmid pLTV1 (containing Tn917-lac) to generate Tn917 transposon mutants. Through the use of temperature manipulation and antibiotic selection, staphylococcal strains harboring this plasmid can be effectively mutagenized to create random libraries amenable to subsequent phenotypic screening and identification of transposon insertion sites.

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Authors and Affiliations

  1. Department of Microbiology and Cell Science, IFAS, University of Florida, Gainesville, FL, 32611-0700, USA

    Kelly C. Rice

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  1. Kelly C. Rice
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Correspondence to Kelly C. Rice .

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Editors and Affiliations

  1. University of Kansas Medical Center, Kansas City, Kansas, USA

    Jeffrey L. Bose

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Rice, K.C. (2014). Creation of Staphylococcal Mutant Libraries Using Transposon Tn917. In: Bose, J. (eds) The Genetic Manipulation of Staphylococci. Methods in Molecular Biology, vol 1373. Humana Press, New York, NY. https://doi.org/10.1007/7651_2014_188

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  • DOI: https://doi.org/10.1007/7651_2014_188

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3157-6

  • Online ISBN: 978-1-4939-3158-3

  • eBook Packages: Springer Protocols

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