Multiplex Genome Editing in Escherichia coli

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Abstract

Lambda Red recombineering is an easy and efficient method for generating genetic modifications in Escherichia coli. For gene deletions, lambda Red recombineering is combined with the use of selectable markers, which are removed through the action of, e.g., flippase (Flp) recombinase. This PCR-based engineering method has also been applied to a number of other bacteria. In this chapter, we describe a recently developed one plasmid-based method as well as the use of a strain with genomically integrated recombineering genes, which significantly speeds up the engineering of strains with multiple genomic alterations.
Original languageEnglish
Title of host publicationSynthetic Metabolic Pathways
Volume1671
Publication date2018
Pages119-129
ISBN (Print)978-1-4939-7294-4
ISBN (Electronic)978-1-4939-7295-1
DOIs
Publication statusPublished - 2018
SeriesMethods in Molecular Biology
ISSN1064-3745

Keywords

  • Flippase recombinase
  • Genome editing
  • Industrial biotechnology
  • Lambda Red recombineering
  • Synthetic biology
  • pSIJ8

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