Generation of Nonpolar Deletion Mutants in Listeria monocytogenes Using the “SOEing” Method

  • Kathrin RychliEmail author
  • Caitriona M. Guinane
  • Karen Daly
  • Colin Hill
  • Paul D. Cotter
Part of the Methods in Molecular Biology book series (MIMB, volume 1157)


The ability to manipulate chromosomally encoded genes is a fundamental biological tool for the analysis of gene function. Here, we provide in greater depth protocols for the creation of nonpolar unlabeled gene deletions in Listeria (L.) monocytogenes that are facilitated by the splicing overlap extension PCR technique. For mutagenesis in L. monocytogenes, we describe two different plasmid-based approaches, which facilitate the introduction of this spliced amplicon in place of the corresponding segment of chromosomal DNA: the pKSV7 system and the pORI280/pVE6007 system.


Nonpolar deletion mutants Listeria monocytogenes SOEing method pKSV7 vector pORI280/pVE6007 system 



This work was supported by the EU FOODSEG grant.


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Kathrin Rychli
    • 1
    Email author
  • Caitriona M. Guinane
    • 2
  • Karen Daly
    • 3
  • Colin Hill
    • 3
    • 4
  • Paul D. Cotter
    • 2
    • 4
  1. 1.University of Veterinary Medicine ViennaViennaAustria
  2. 2.Teagasc Food Research Centre, MooreparkFermoy, County CorkIreland
  3. 3.Department of MicrobiologyUniversity College CorkCorkIreland
  4. 4.Alimentary Pharmabiotic CentreUniversity College CorkCorkIreland

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