Open AccessTransposon Tn5 mutagenesis of Pseudomonas fluorescens to isolate mutants deficient in antibacterial activity
Author(s) -
Rajendran Narayanan,
Jahn Dieter,
Jayaraman Kunthala,
Marahiel Mohamed A.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb06636.x
Subject(s) - pseudomonas fluorescens , transposable element , transposon mutagenesis , mutagenesis , mutant , biology , insertional mutagenesis , auxotrophy , plasmid , genetics , sleeping beauty transposon system , pseudomonas , insertion , microbiology and biotechnology , gene , bacteria
Pseudomonas fluorescens was subjected to insertion mutagenesis studies using the transposon Tn5‐GM to generate mutants deficient in antibacterial activity minus mutants. The transposon located on the temperature‐sensitive plasmid pCHR84 was conjugally transferred into the non‐pathogenic pseudomonad using the triparental mating procedure. Random integration of Tn 5 ‐GM into the chromosome of P. fluorescens was achieved by heat ttreatment of the transformed cells at 42°C. Approximately 2% of transconjugants revealed an auxotrophic phenotype indicating efficient integration of the employed transposon into the chromosome of P. fluorescens . One transposon insertion mutant was obtained showing an antibacterial activity minus phenotype. This mutant (MM‐7) was found to be defective in the production of an unidentified antibacterial compound against B. subtilis . These results introduce Tn 5 transposon mutagenesis as a new useful tool for the molecular analysis of P. fluorescens .