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

Kathrin Rychli; Caitriona M. Guinane; Karen Daly; Colin Hill; Paul D. Cotter

doi:10.1007/978-1-4939-0703-8_16

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

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

Research output: Contribution to journal › Article › peer-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 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.

Original language	English
Pages (from-to)	187-200
Number of pages	14
Journal	Methods in Molecular Biology
Volume	1157
DOIs	https://doi.org/10.1007/978-1-4939-0703-8_16
Publication status	Published - 2014

Keywords

Listeria monocytogenes
Nonpolar deletion mutants
PKSV7 vector
PORI280/pVE6007 system
SOEing method

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10.1007/978-1-4939-0703-8_16

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title = "Generation of nonpolar deletion mutants in listeria monocytogenes using the “SOEing” method",

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 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.",

keywords = "Listeria monocytogenes, Nonpolar deletion mutants, PKSV7 vector, PORI280/pVE6007 system, SOEing method",

author = "Kathrin Rychli and Guinane, \{Caitriona M.\} and Karen Daly and Colin Hill and Cotter, \{Paul D.\}",

note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media New York 2014.",

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doi = "10.1007/978-1-4939-0703-8\_16",

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AU - Rychli, Kathrin

AU - Guinane, Caitriona M.

AU - Daly, Karen

AU - Hill, Colin

AU - Cotter, Paul D.

N1 - Publisher Copyright: © Springer Science+Business Media New York 2014.

PY - 2014

Y1 - 2014

N2 - 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.

AB - 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.

KW - Listeria monocytogenes

KW - Nonpolar deletion mutants

KW - PKSV7 vector

KW - PORI280/pVE6007 system

KW - SOEing method

UR - https://www.scopus.com/pages/publications/84922022670

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DO - 10.1007/978-1-4939-0703-8_16

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AN - SCOPUS:84922022670

SN - 1064-3745

VL - 1157

SP - 187

EP - 200

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

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

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