Table 1.

Strains and plasmids used in this study

Strain or plasmidGenotype or characteristicsaSource or reference
E. coli
  DH5αFφ80dlacZΔM15 recA1 endA1 gyrA96 thi-1 relA1 supE44 hsdR17(rK mK+) Δ(lacZYA-argF)U169Gibco-BRL
  SM10Mobilizer strain47
  BL21(λDE3)FompT hsdSB(rB mB) gal dcm (DE3)Novagen
  CSH7lacY rpsL thi-131
  UM1As CSH7 pluskatE1 katG1431
P. aeruginosa
  PAO1Wild type; prototroph22
  PAO1katAGmrkatA::GmThis study
  PAO1bfrAGmrΔbfrA::GmThis study
  PAO1 katA bfrAGmr ΔkatA bfrA::GmThis study
  FRD2katBalgT18 katB::Gm10
 pBluescript(KS)- or pBluescript (KS)+Extended polylinker pUC derivativeStratagene
 pKS-TAApr; TA PCR cloning vector that uses EcoRV site for cloning purposesThis study
 pCRIIApr; TA PCR cloning vectorInVitrogen
 pCR2.1Apr; TA PCR cloning vectorInVitrogen
 pUCGMApr Gmr; pUC19 + 850-bp Gmr cassette45
 pPZ30Apr; broad-host-range lacZ-based promoter probe vector43
 pUCP19Apr; broad-host-range expression vector52
 pUCP21TApr; broad-host-range expression vector52
 pEX100TApr Cbr; mobilizableoriT sacB vector for construction of mutants46
 pRK2013Kmr Ori (ColE1) OriT (Mob+) Tra+14
 pET23aApr; overexpression vectorNovagen
 pJFM12Apr; pKS− with 3.6-kbEcoRI-EcoRV fragment containing rpsD′-rpoA-rplQ-katA-bfrAThis study
 pJFM13Apr; pEX100T with blunted 3.6-kbEcoRI-EcoRV fragment containingrpsD′-rpoA-rplQ-katA-bfrA′ in the SmaI site of the vectorThis study
 pJFM14AprGmr; pJFM13 with 850-bp aaC1 cassette within theSmaI site of katAThis study
 pJFM15Apr Gmr; pEX100T with bluntedEcoRI fragment from pBFR1020ΔbfrA::Gm and with 850-bp aaC1 cassette within the deleted 520-bpNdeI-SstII fragment, deletingbfrAThis study
 pJFM16AprGmr; pJFM13 with 850-bp aaC1 cassette within the deleted 1,311-bp SmaI-NdeI site, deletingkatA and bfrAThis study
 pJFM17Apr; pKS− with 1,450-bp katAPCR productThis study
 pJFM18Apr; pET23a withNcoI-EagI katAThis study
 pRP411Apr; pKS+ containing 411-bpSalI-EcoRV fragment of katA-bfrAintergenic regionThis study
 pBFR1020pCR2.1 harboring 1,020-bp PCR fragment containing bfrAThis study
 pBFR1020ΔbfrA::GmpBFR1020 with 520-bpNdeI-SstII deletion of bfrA replaced by aaC1 cassetteThis study
 pPZ-katAApr; pPZ30 containing thekatA promoter on a 756-bp EcoRI-PstI fragment translationally fused to lacZThis study
 pPZ-bfrAApr; pPZ30 containing thebfrA promoter on a 389-bp EcoRI-PstI fragment translationally fused to lacZThis study
 pBFR4pUCP19 with wild-type bfrA geneThis study
 pBFR18pUCP19 with bfrA18 encoding Bfr18 (E18K)This study
 PBFR25pUCP19 with bfrA25encoding Bfr25 (Y25I)This study
  • a Abbreviations used for genetic markers were those described by Holloway et al. (22). Mob+, mobilization site (ColE1); Tra+, conjugative phenotype; oriT, origin of transfer (RK2); Apr, ampicillin resistance; Cmr, chloramphenicol resistance; TA, thymine-adenine; pKS−, pBluescript KS(−); pKS+, pBluescript (KS)+.