Table 1

Bacterial strains and plasmids used in this study

Strain or plasmidDescriptionReference or source
Strains
    PAO1P. aeruginosa PAO1 WTa41
    PAO1 ampC::FRTWT with FRT scar at nucleotide 1070 of ampC (PA4110)This study
    PAO1 dacB::FRTWT with FRT scar at nucleotide 168 of dacB (PBP4, PA3047)16
    PAO1 sltB1::FRTWT with FRT scar at nucleotide 577 of sltB1 (PA4001)This study
    PAO1 mgtA::TnWT with transposon ISlacZ/hah insertion at nucleotide 495 of mgtA (PA0378)41
    PAO1 pbpG::TnWT with transposon ISlacZ/hah insertion at nucleotide 282 of pbpG (PBP7, PA0869)41
    PAO1 sltG::TnWT with transposon ISphoA/hah insertion at nucleotide 227 of sltG (PA1171)41
    PAO1 mltA::TnWT with transposon ISphoA/hah insertion at nucleotide 397 of mltA (PA1222)41
    PAO1 mltD::TnWT with transposon ISphoA/hah insertion at nucleotide 868 of mltD (PA1812)41
    PAO1 pbpC::TnWT with transposon ISphoA/hah insertion at nucleotide 1226 of pbpC (PBP3a, PA2272)41
    PAO1 mltF2::TnWT with transposon ISlacZ/hah insertion at nucleotide 991 of mltF2 (PA2865)41
    PAO1 slt::TnWT with transposon ISlacZ/hah insertion at nucleotide 1256 of slt (PA3020)41
    PAO1 mltF::TnWT with transposon ISphoA/hah insertion at nucleotide 730 of mltF (PA3764)41
    PAO1 sltH::TnWT with transposon ISlacZ/hah insertion at nucleotide 415 of sltH (PA3992)41
    PAO1 dacC::TnWT with transposon ISlacZ/hah insertion at nucleotide 549 of dacC (PBP6, PA3999)41
    PAO1 mltB::TnWT with transposon ISphoA/hah insertion at nucleotide 358 of mltB (PA4444)41
    PAO1 ponA::TnWT with transposon ISphoA/hah insertion at nucleotide 1573 of ponA (PBP1a, PA5045)41
    PAO1 mrcB::TnWT with transposon ISphoA/hah insertion at nucleotide 973 of mrcB (PBP1b, PA4700)41
    PAO1 sltB1::FRT dacB::FRTsltB1::FRT mutant with FRT scar at nucleotide 168 of dacB (PBP4)This study
    PAO1 mltB::Tn dacB::FRTmltB::Tn mutant with FRT scar at nucleotide 168 of dacB (PBP4)This study
    PAO1 mltB::Tn sltB1::FRTmltB::Tn mutant with FRT scar at nucleotide 577 of sltB1This study
    PAO1 ampC::FRT dacB::FRTampC::FRT with FRT scar at nucleotide 168 of dacB (PBP4)This study
    PAO1 sltB1::FRT ampC::FRTsltB1::FRT mutant with FRT scar at nucleotide 1070 of ampCThis study
    PAO1 mltB::Tn ampC::FRTmltB::Tn mutant with FRT scar at nucleotide 1070 of ampCThis study
    PAO1 pBADGr-dacBWT with pBADGr derivative containing dacBThis study
    PAO1 pBADGr-sltB1WT with pBADGr derivative containing sltB1This study
    PAO1 dacB::FRT + pBADGr-dacBdacB::FRT mutant with pBADGr derivative containing dacBThis study
    PAO1 sltB1::FRT + pBADGr-sltB1sltB1::FRT mutant with pBADGr derivative containing sltB1This study
    PAO1 sltB1::FRT dacB::FRT + pBADGr-dacBsltB1::FRT dacB::FRT mutant with pBADGr derivative containing dacBThis study
    PAO1 mltB::Tn dacB::FRT + pBADGr-dacBmltB::Tn dacB::FRT mutant with pBADGr derivative containing dacBThis study
    PAO1 sltB1::FRT dacB::FRT + pBADGr-sltB1sltB1::FRT dacB::FRT mutant with pBADGr derivative containing sltB1This study
    PAO1 mltB::Tn/sltB1::FRT + pBADGr-sltB1mltB::Tn sltB1::FRT mutant with pBADGr derivative containing sltB1This study
Plasmids
    pPS856Source of FRT-flanked gentamicin cassette; Gmr12
    pEX18ApSuicide vector used for gene replacement; Apr12
    pFLP2Suicide vector encoding Flp recombinase; Apr12
    pBADGrpMLBAD backbone with dhfr replaced with aacC1; Gmr42
    pEX18Ap-dacB::FRTGmFRTSuicide vector containing dacB disrupted at position 168 with FRT-flanked gentamicin resistance cassette; Apr Gmr16
    pEX18Ap-sltB1::FRTGmFRTSuicide vector containing sltB1 disrupted at position 577 with FRT-flanked gentamicin resistance cassette; Apr GmrThis study
    pEX18Ap-ampC::FRTGmFRTSuicide vector containing ampC disrupted at position 1070 with FRT-flanked gentamicin resistance cassette; Apr GmrThis study
    pBADGr-dacBpBADGr derivative containing dacB on an EcoRI fragment; GmrThis study
    pBADGr-sltB1pBADGr derivative containing sltB1 on an EcoRI fragment; GmrThis study
  • a WT, wild type.