A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Generation of Single-Copy Transposon Insertions in Clostridium perfringens by Electroporation of Phage Mu DNA Transposition Complexes
Tekijät: Lanckriet A, Timbermont L, Happonen LJ, Pajunen MI, Pasmans F, Haesebrouck F, Ducatelle R, Savilahti H, Van Immerseel F
Kustantaja: AMER SOC MICROBIOLOGY
Julkaisuvuosi: 2009
Lehti:: Applied and Environmental Microbiology
Tietokannassa oleva lehden nimi: APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Lehden akronyymi: APPL ENVIRON MICROB
Vuosikerta: 75
Numero: 9
Aloitussivu: 2638
Lopetussivu: 2642
Sivujen määrä: 5
ISSN: 0099-2240
DOI: https://doi.org/10.1128/AEM.02214-08
Tiivistelmä
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.
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.
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