Generation of Single-Copy Transposon Insertions in Clostridium perfringens by Electroporation of Phage Mu DNA Transposition Complexes
| Autor: | Frank Pasmans, Maria Pajunen, Freddy Haesebrouck, Richard Ducatelle, Anouk Lanckriet, Harri Savilahti, Lotta Happonen, F. Van Immerseel, Leen Timbermont |
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| Rok vydání: | 2009 |
| Předmět: |
DNA
Bacterial Transposable element Tn3 transposon Clostridium perfringens Mutagenesis (molecular biology technique) Genetics and Molecular Biology Biology medicine.disease_cause Applied Microbiology and Biotechnology Bacteriophage mu Insertional mutagenesis 03 medical and health sciences medicine Gene Library 030304 developmental biology Recombination Genetic Genetics 0303 health sciences Ecology 030306 microbiology Sleeping Beauty transposon system Mutagenesis Insertional Electroporation Transposon mutagenesis Bacteriophage Mu Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 75:2638-2642 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.02214-08 |
| Popis: | 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 Tn 916 -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 . |
| Databáze: | OpenAIRE |
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