TABLE 1.

Bacterial strains, plasmids, and oligonucleotides used for this study

Strain, plasmid, or oligonucleotideRelevant characteristics or sequence (5′-3′)Source or reference
Strains
    P. aeruginosa PAO1Prototroph, nonmucoidATCC 15692
    P. aeruginosa PAO1ΔpslAPAO1 ΔpslA mutant, biofilm negativeThis study
    E. coli S17-1recA; harbors the tra genes of plasmid RP4 in the chromosome; proA thi-135
    E. coli XL1-BluerecA1 endA1 gyrA96 thi1 hsdR17 (rK mK+) supE44 relA1 λlac [F′ proAB lacIqZΔM15 Tn10(Tc)]3
Plasmids
pBluescript SK(−)Ampr lacPOZ, T7 and T3 promoterStratagene
pEX100TAmpr oriT sacB lacPOZ′34
pGEM-TeasyAmpr lacPOZ′Promega
pPS856Ampr Gmr FRT17
pEXpslApEX100T containing GM cassette flanked by 5′ and 3′ ends of the pslA gene
pTZ110Broad-host-range lacZ fusion vector33
pBBR1-MCS5Gmr lacPOZmob, broad-host-range vector20
pBBR1-MCS5::pslApBBR1-MCS5 containing EcoRI/BamHI PCR fragment of pslAThis study
pSKPpslpBluescript SK(−) containing EcoRI/BamHI PCR fragment of psl promoterThis study
pTZ110::PpslpTZ110 containing EcoRI/BamHI PCR fragment of psl promoterThis study
pProbeAT′Broad-host-range gfp (stable form) fusion vector24
pProbeAT′::PpslpProbeAT′ containing EcoRI/BamHI PCR fragment of psl promoterThis study
pProbe′-gfp[AAV]Broad-host-range gfp (unstable form) fusion vector
pProbe′-gfp[AAV]::PpslpProbe′-gfp[AAV] containing EcoRI/BamHI PCR fragment of psl promoter24
pProbe′-gfp[AAV]::ΩGm-PpslpProbe′-gfp[AAV]::Ppsl containing a gentamicin restistance gene cassette in the opposite direction to that of the gfp geneThis study
pGEM-Teasy::RACEpGEM-Teasy containing 5′-RACE PCR productThis study
Oligonucleotides
PA2231-KO-UAGTAGATAGCCTGTCGCTGACACGGGThis study
PA2231-KO-DAAAAAAGGATCCGATCACCGTCCGTTGCAGGTACATGThis study
PA2231-KO-UBAAAAAGGATCCCGGCTGATCTGGGTCTTCAAGTTCCThis study
PA2231-KO-DBTCAGTAGACTTCCTTGGTCAGGAGThis study
PA2231-UEAAAAAGAATTCTGGACTGCCCGTGATCGGCAGAGCThis study
PA2231-DBAAAAAGGATCCGCAGGGGATCAGCGGGGCGACGGCThis study
PpslUEAAAAAGAATTCCCGAAATGGCACGAGGCGGCGCTGThis study
PpslDBAAAAAGGATCCGCCGATCACGGGCAGTCCATTGAThis study
RACE-primer1CAGGCCCAGGCCCTGGTACATGAACAACAGCAGGCThis study
RACE-primer2CGTCGGAGTAGACGTCCAGTGCCTGGAACThis study
RACE-primer3GCGCAGCTCCACGGGAACCAGGCCGGGGThis study