Genome-scale identification of resistance functions in Pseudomonas aeruginosa using Tn-seq.

ABSTRACT: We describe a deep-sequencing procedure for tracking large numbers of transposon mutants of Pseudomonas aeruginosa. The procedure employs a new Tn-seq methodology based on the generation and amplification of single-strand circles carrying transposon junction sequences (the Tn-seq circle method), a method which can be used with virtually any transposon. The procedure reliably identified more than 100,000 transposon insertions in a single experiment, providing near-saturation coverage of the genome. To test the effectiveness of the procedure for mutant identification, we screened for mutations reducing intrinsic resistance to the aminoglycoside antibiotic tobramycin. Intrinsic tobramycin resistance had been previously analyzed at genome scale using mutant-by-mutant screening and thus provide

SUBMITTER: Gallagher LA 

PROVIDER: S-EPMC3023915 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature