Generation of Markerless Gene Deletion Mutants in Listeria monocytogenes Using a Mutated pheS for Counterselection

Shai Ran Sapir, Etai Boichis, Anat A. Herskovits*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Gene alteration/deletion by allelic exchange is the preferred strategy for gene manipulation in bacteria. Here we present the fundamentals for an efficient allelic exchange gene deletion method in the bacterial pathogen Listeria monocytogenes. Combining vector generation by Gibson assembly with a counterselection system based on the mutated phenylalanine synthetase (pheS*) makes the generation of gene deletion mutants straightforward and time efficient.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages3-10
Number of pages8
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2427
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Allelic exchange
  • Counterselection
  • Gibson assembly
  • Listeria monocytogenes
  • Mutagenesis
  • pheS

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