Article

Transposition of Tn5367 in Mycobacterium marinum, Using a Conditionally Recombinant Mycobacteriophage

University of Cologne, Köln, North Rhine-Westphalia, Germany
Journal of Bacteriology (Impact Factor: 2.81). 04/2003; 185(5):1745-8. DOI: 10.1128/JB.185.5.1745-1748.2003
Source: PubMed

ABSTRACT

Mycobacterium marinum is a close relative of the obligate human pathogen Mycobacterium tuberculosis. As with M. tuberculosis, M. marinum causes intracellular infection of poikilothermic vertebrates and skin infection in humans. It is considered a valid model
organism for the study of intracellular pathogenesis of mycobacteria. Low transformation efficiencies for this species have
precluded approaches using mutant libraries in pathogenesis studies. We have adapted the conditionally replicating mycobacteriophage
phAE94, originally developed as a transposon mutagenesis tool for M. tuberculosis, to meet the specific requirements of M. marinum. Conditions permissive for phage replication in M. tuberculosis facilitated highly efficient transposon delivery in M. marinum. Using this technique we succeeded in generating a representative mutant library of this species, and we conclude that TM4-derived
mycobacteriophages are temperature-independent suicide vectors for M. marinum.

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    • ") carrying the kanamycin resistance transposon Tn5367 ( Shin et al . , 2006 ) was propagated in M . smegmatis mc 2 155 ( Bardarov et al . , 1997 ) and used to infect M . marinum as described previously ( Rybniker et al . , 2003 ) ."
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