Table 1.

Strains and plasmids

Strain or plasmidGenotype and/or descriptionReference or source
P. aeruginosa
  8830his-1 ppk+9
  8830K::Tchis-1 ppk mutantThis study
E. coli
  DH5αFendA1 hsdR17 supE44 thi-1 recA1 gyrA relA1 ΔlacU169φ80lacZΔM1527
  BL21(DE3)(pLysS)FompT hsdSB (rBmB) gal dcm (λDE3) pLysSNovagen
 pCR2.1AmprKmr; vector for TA cloning of PCR productsInVitrogen
 pBR322Ampr Tcr; E. coliplasmid cloning vector7
 pUC19Ampr; E. coli plasmid cloning vector39
 pNOT19pUC19 derivative; 10-nucleotide NotI oligonucleotide inserted intoNdeI site33
 pMOB2Kmr; mobilization cassette33
 pET24a(+)Kmr; T7lac promoter, His · Tag C-terminal fusionNovagen
 pET28a(+)Kmr; T7lacpromoter, His · Tag N-terminal fusionNovagen
 pQF50Ampr; broad-host-range promoter probe vector11
 pBBR1MCSCmr; broad-host-range plasmid cloning vector20
 pCR-F2R51.7-kb PCR probe cloned in pCR2.1This study
 pPPKB3.5-kb SphI fragment containing theppk gene cloned into pBR322This study
 pPPX53-kb SalI fragment containing theppx gene cloned into pUC19This study
 pPPK-Hisppk PCR-generated cloned intoEcoRI and HindIII sites of pET24a(+)This study
 pPPX-Hisppx PCR-generated cloned intoNdeI and HindIII sites of pET28a(+)This study
 pNotK::TcpNot19 derivative, 1.6-kbKpnI-HindIII PCR fragment and Tcrcassette in BamHI siteThis study
 pQF-376K376-bp XhoI fragment containing theppk promoter cloned into SalI site of pQF50This study
 pQF-649X649-bp SphI fragment containing the putative ppx promoter cloned into pQF50This study
 pCR-52,488-bp PCR fragment containing theppk gene and its promoter cloned into pCR2.1This study
 pBBR-KHindIII-EcoRV fragment derived from pCR-5 cloned into pBBR1MCSThis study