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Lambda Red–Mediated Recombination in Shiga Toxin-Producing Escherichia coli

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Shiga Toxin-Producing E. coli

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2291))

Abstract

The bacteriophage Lambda (λ) “Red” recombination system has enabled the development of efficient methods for engineering bacterial chromosomes. This system has been particularly important to the field of bacterial pathogenesis, where it has advanced the study of virulence factors from Shiga toxin-producing and enteropathogenic Escherichia coli (STEC and EPEC). Transient plasmid-driven expression of Lambda Red allows homologous recombination between PCR-derived linear DNA substrates and target loci in the STEC/EPEC chromosomes. Red-associated techniques can be used to create individual gene knockouts, generate deletions of large pathogenicity islands, and make markerless allelic exchanges. This chapter describes specific strategies and procedures for performing Lambda Red-mediated genome engineering in STEC.

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Correspondence to Kenneth G. Campellone .

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Campellone, K.G., Coulter, A.M. (2021). Lambda Red–Mediated Recombination in Shiga Toxin-Producing Escherichia coli. In: Schüller, S., Bielaszewska, M. (eds) Shiga Toxin-Producing E. coli . Methods in Molecular Biology, vol 2291. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1339-9_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1339-9_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1338-2

  • Online ISBN: 978-1-0716-1339-9

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