Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant genotype or phenotypeReference or source
Strains
P. aeruginosa
  PAO1Wild typeHolloway collection
  PANO67NTGa mutant derived from PAO1 34
  PAO1lecA::lux lecA::luxCDABEgenomic reporter fusion in PAO1This study
  PAOR lasR mutant derived from PAO1 38
  PDO100 rhlI mutant derived from PAO1 6
  PDO111 rhlR mutant derived from PAO1 6
  PAO1 rpoSnegative rpoS mutant derived from PAO1 35
E. coli
  JM109 recA1 supE44 endA1 hsdR17 gyrA96 relA1 thiΔ(lac-proAB) F′ [traD36 proAB+lacI q lacZΔM15] 72
  S17-1 λpir thi pro hsdR hsdM+ recARP4-2-Tc::Mu-Km::Tn7 λpir 61
  CC118 λpirΔ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rps-1 rpoB argE(Amp) recA thi pro hsdRM + RP4-2-Tc::Mu-Km::Tn7 λpir 32
  XL1-Blue MRΔ(mcrA)183Δ(mcrCB-hsdMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac Stratagene
Plasmids
 pSB421 EcoRI fragment containingluxCDABE cassette and Kmr element cloned into pUC18Not 70
 pUCP18Like pUC18 but additional 1.8-kb stabilizing fragment for maintenance inPseudomonas spp. 60
 pKNG101Suicide vector carrying the sacBR genes for sucrose sensitivity; Smr 36
 pNQluxIII luxCDABE genes cloned into pNQ705This laboratory
 pNQ705Cmr suicide plasmid derived from pGP704 45
 Supercos1Cosmid vector, pBR322-derived ori, Ampr Stratagene
 pMW47.12-kb PstI PAO1 DNA insert (rhlRI) in pUCP18 37
 pMW471.21.3-kb PstI-EcoRI fragment containing rhlR in pUCP18 37
 plasR lasR locus cloned into pUCP18This laboratory
 pCF6b32.6-kb PAO1 DNA insert containing the lecA gene in Supercos1This study
 pCF14.4-kb PstI fragment derived from pCF6b containing the lecA gene in pUCP18This study
 pCF23.1-kb PstI-HindIII fragment derived from pCF6b containing the lecA gene in pUCP18This study
 pCF3600-bp EcoRI fragment derived from pCF6b containing the lecA gene in pUCP18This study
 pCol2PCR fragments encoding thelecA gene region cloned into EcoRI andKpnI sites upstream and downstream ofluxCDABE Kmr in pSB421This study
 pCol4∼8-kb NotI fragment from pCol2 cloned into pKNG101This study
 pCol9496-bp PCR fragment, encoding thelecA upstream region, cloned into the NotI site of pNQluxIIIThis study
  • a NTG,N-methyl-N′-nitro-N-nitrosoguanidine.