Generation of nonpolar deletion mutants in listeria monocytogenes using the “soeing” method

  • Kathrin Rychli
  • , Eva Wagner
  • , Caitriona M. Guinane
  • , Karen Daly
  • , Colin Hill
  • , Paul D. Cotter

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

Abstract

The ability to manipulate chromosomally encoded genes is a fundamental biological tool for the analysis of gene function. Here, we provide in greater depth a protocol for the creation of nonpolar unlabelled gene deletions in Listeria monocytogenes that are facilitated by the splicing overlap extension PCR technique. For mutagenesis in L. monocytogenes, we describe the pKSV7 plasmid-based approach, which facilitates the introduction of a spliced amplicon in place of the corresponding segment of chromosomal DNA.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages165-175
Number of pages11
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2220
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Listeria monocytogenes
  • Nonpolar deletion mutants
  • pKSV7 vector
  • SOEing method

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