TABLE 1.

Strains, plasmids, and primers

Strain, plasmid, or primerGenotype, characteristics, or sequenceaReference or source
Strains
    P. aeruginosa PAO158
    E. coli JM105Pharmacia
    E. coli cc118 λ-pirΔ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoE(Am) recA1; lysogenized with Δpir phage18
    E. coli s17 λ-pirthi pro hsdR recA RP4-2 (Tet::Mu) (Km::Tn7) λ56
    LM118PAO1 ΔmutY::GmThis study
    LM117PAO1 ΔmutT::GmThis study
    LM121PAO1 ΔmutM::GmThis study
    LM76PAO1 ΔmutY::Gm with pLM100This study
    LM79PAO1 ΔmutT::Gm with pLM101This study
    LM82PAO1 ΔmutM::Gm with pLM102This study
Plasmids
    pUC18notApr; identical to pUC18 but with NotI/polylinker of pUC18/NotI as MCS18
    pCK318RP4mob oriR6K sacB bla16
    pUCP26P. aeruginosa-E. coli shuttle vector62
    pLM100pUCP26 containing 1.2-kb BamHI-HindIII PCR fragment of PAO1 mutYThis study
    pLM101pUCP26 containing 1-kb BamHI-HindIII PCR fragment of PAO1 mutTThis study
    pLM102pUCP26 containing 0.9-kb XbaI-HindIII PCR fragment of PAO1 mutMThis study
    pLM103pUCP26 containing 1.95-kb BamHI-HindIII PCR fragment of PAO1 mutLThis study
Primers for PCR
    mutY-fw-tjek5′-GACAAGGAAGGCATGGGCAAGGTC-3′This study
    mutY-rev-tjek5′-GGTACTTGCGGCACATCACGGTG-3′This study
    mutM-fw-tjek5′-CGACTACGACTTCGAGAACCTCAAGG-3′This study
    mutM-rev-tjek5′-GGACATCGTGCACCTTTCTTATGGCAG-3′This study
    mutT-fw-tjek5′-TATGTCGAGACGATTTATCAGTGGCCTG-3′This study
    mutT-rev-tjek5′-GACCAGGAGGAAACGCTGGAGG-3′This study
    nfxB fw 15′-GCACAATGCGCACAATCAG-3′This study
    nfxB rev 15′-TCGGTCCGTGCCATGC-3′This study
Primers for complementation
    mutY fw m. BamHI+SD5′-CGCGGATCCGCGAGGAGAAGAGCTA CGCAAATGACACCTGAAGGC-3′This study
    mutY rev m. HindIII5′-CCCAAGCTTGGGTCATGGTCGTTCTCCTGC-3′This study
    mutM fw m. XbaI+SD5′-GCTCTAGAGCAGGAGAAGTGCAATGCG CATGCCCGAACTACCCGAAG-3′This study
    mutM rev m. HindIII5′-CCCAAGCTTGGGCTTCTTGTGGCGGTAGAGTATGC-3′This study
    mutT fw m. BamHI+SD5′-CGCGGATCCGCGAGGAGAAGTGCAATGC CCGTGAAACGAGTACATGTC-3′This study
    mutT rev m. HindIII5′-CCCAAGCTTGGG TTCATTCCACCGTCAAAGGC-3′This study
    mutL fw. m. BamHI+SD5′-CGCGGATCCGCG AGGAGAAG TTCA CCAGTGATGAGTGAAGCACC-3′This study
    mutL rev m. HindIII5′-CCCAAGCTTGGG GAAGCTGCAAGGCTCAGA-3′This study
Primers for RT-PCR
    MexF-L RTpcr5′-TCTACGACCCGACCATCTTC-3′This study
    MexF-R RTpcr5′-CAGGTCTGCAGGAACAGGAT-3′This study
    MexY-L RTpcr5′-GCCCAACGACATCTACTTCAA-3′This study
    MexY-R RTpcr5′-CATGCCTTCCTGGTAATGGT-3′This study
    MexD-L RTpcr5′-TCAACGGTCTGGGTAACTCC-3′This study
    MexD-R RTpcr5′-TGGATCTCGCCAAGAAGAGT-3′This study
    MexB-L RTpcr5′-TACGAAAGCTGGTCGATTCC-3′This study
    MexB-R RTpcr5′-GAAGAACACGTCGTTGGACA-3′This study
  • a MCS, multiple cloning site. Underlined nucleotides in sequences of primers for complementation are Shine-Dalgarno motifs.