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Mutator gene analysis. BLASTing the E. coli mutator gene sequences (TIGR) against the database of PAO1 (Pseudomonas Genome Project) allowed us to obtain the corresponding putative sequences of P. aeruginosa mutator genes: mutS, mutL, mutT, mutY, mutM, mutD, and uvrD. Specific primers were designed to amplify these genes in the studied strains by PCR.
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DNA from these four isolates was checked by amplifying in the same PCR tube another gene with higher molecular weight (uvrD) that, in contrast to mutY, was correctly amplified.
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2 decrease in mutation frequency) was found in mutator strains from 4 out of the 11 CF patients with mutators. No significant change in mutation frequency was detectable in non-mutator strains or with the plasmid lacking the mutS gene.
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We thank B. Levin, J. M. Gómez-Gómez, I. Matic, and J. L. Martinez for useful discussions and comments, and two anonymous referees for suggestions; H. Escobar (Unit of Cystic Fibrosis) for continuous support; and R. Garber (PathoGenesis) for BLASTing the E. coli mutS sequence against the database of PAO1. We also thank L. de Rafael for English correction, O. Santisteban for help with preparation of the figures, and A. Vindel (National Center for Microbiology, Carlos III Institute, Majadahonda, Madrid) for help with serotyping and phagetyping of Pseudomonas isolates. Sequence data for this work were obtained from The Institute for Genomic Research (TIGR) and the Pseudomonas Genome Project.
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