Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes

A. Lanckriet, L. Timbermont, Lotta Johanna Happonen, Maria Pajunen, F. Pasmans, F. Haesebrouck, R. Ducatelle, Harri Savilahti, F. Van Immerseel

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was successfully used to generate a single-insertion mutant library both for a laboratory strain and for a field isolate. Thus, it can be used as a tool in large-scale screening to identify virulence genes of C. perfringens.
Original languageEnglish
JournalApplied and Environmental Microbiology
Volume75
Issue number9
Pages (from-to)2638-2642
Number of pages5
ISSN0099-2240
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fields of Science

  • GRAM-POSITIVE BACTERIA
  • ESCHERICHIA-COLI
  • ENTEROCOCCUS-FAECALIS
  • CONJUGATIVE TRANSPOSITION
  • MYCOPLASMA-GALLISEPTICUM
  • TN916
  • MUTAGENESIS
  • IDENTIFICATION
  • GENES
  • EFFICIENT
  • 118 Biological sciences

Cite this

Lanckriet, A. ; Timbermont, L. ; Happonen, Lotta Johanna ; Pajunen, Maria ; Pasmans, F. ; Haesebrouck, F. ; Ducatelle, R. ; Savilahti, Harri ; Van Immerseel, F. / Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes. In: Applied and Environmental Microbiology. 2009 ; Vol. 75, No. 9. pp. 2638-2642.
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title = "Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes",
abstract = "Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was successfully used to generate a single-insertion mutant library both for a laboratory strain and for a field isolate. Thus, it can be used as a tool in large-scale screening to identify virulence genes of C. perfringens.",
keywords = "GRAM-POSITIVE BACTERIA, ESCHERICHIA-COLI, ENTEROCOCCUS-FAECALIS, CONJUGATIVE TRANSPOSITION, MYCOPLASMA-GALLISEPTICUM, TN916, MUTAGENESIS, IDENTIFICATION, GENES, EFFICIENT, 118 Biological sciences",
author = "A. Lanckriet and L. Timbermont and Happonen, {Lotta Johanna} and Maria Pajunen and F. Pasmans and F. Haesebrouck and R. Ducatelle and Harri Savilahti and {Van Immerseel}, F.",
year = "2009",
doi = "10.1128/AEM.02214-08",
language = "English",
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Lanckriet, A, Timbermont, L, Happonen, LJ, Pajunen, M, Pasmans, F, Haesebrouck, F, Ducatelle, R, Savilahti, H & Van Immerseel, F 2009, 'Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes' Applied and Environmental Microbiology, vol. 75, no. 9, pp. 2638-2642. https://doi.org/10.1128/AEM.02214-08

Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes. / Lanckriet, A.; Timbermont, L.; Happonen, Lotta Johanna; Pajunen, Maria; Pasmans, F.; Haesebrouck, F.; Ducatelle, R.; Savilahti, Harri; Van Immerseel, F.

In: Applied and Environmental Microbiology, Vol. 75, No. 9, 2009, p. 2638-2642.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes

AU - Lanckriet, A.

AU - Timbermont, L.

AU - Happonen, Lotta Johanna

AU - Pajunen, Maria

AU - Pasmans, F.

AU - Haesebrouck, F.

AU - Ducatelle, R.

AU - Savilahti, Harri

AU - Van Immerseel, F.

PY - 2009

Y1 - 2009

N2 - Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was successfully used to generate a single-insertion mutant library both for a laboratory strain and for a field isolate. Thus, it can be used as a tool in large-scale screening to identify virulence genes of C. perfringens.

AB - Transposon mutagenesis is a tool that is widely used for the identification of genes involved in the virulence of bacteria. Until now, transposon mutagenesis in Clostridium perfringens has been restricted to the use of Tn916-based methods with laboratory reference strains. This system yields primarily multiple transposon insertions in a single genome, thus compromising its use for the identification of virulence genes. The current study describes a new protocol for transposon mutagenesis in C. perfringens, which is based on the bacteriophage Mu transposition system. The protocol was successfully used to generate a single-insertion mutant library both for a laboratory strain and for a field isolate. Thus, it can be used as a tool in large-scale screening to identify virulence genes of C. perfringens.

KW - GRAM-POSITIVE BACTERIA

KW - ESCHERICHIA-COLI

KW - ENTEROCOCCUS-FAECALIS

KW - CONJUGATIVE TRANSPOSITION

KW - MYCOPLASMA-GALLISEPTICUM

KW - TN916

KW - MUTAGENESIS

KW - IDENTIFICATION

KW - GENES

KW - EFFICIENT

KW - 118 Biological sciences

U2 - 10.1128/AEM.02214-08

DO - 10.1128/AEM.02214-08

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SP - 2638

EP - 2642

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

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