Bacterial strains, plasmids, and oligonucleotides used for this study
| Strain, plasmid, or oligonucleotide | Relevant characteristics or sequence (5′-3′) | Source or reference |
|---|---|---|
| Strains | ||
| P. aeruginosa PAO1 | Prototroph, nonmucoid | ATCC 15692 |
| P. aeruginosa PAO1ΔpslA | PAO1 ΔpslA mutant, biofilm negative | This study |
| E. coli S17-1 | recA; harbors the tra genes of plasmid RP4 in the chromosome; proA thi-1 | 35 |
| E. coli XL1-Blue | recA1 endA1 gyrA96 thi1 hsdR17 (rK− mK+) supE44 relA1 λ−lac [F′ proAB lacIqZΔM15 Tn10(Tc)] | 3 |
| Plasmids | ||
| pBluescript SK(−) | Ampr lacPOZ, T7 and T3 promoter | Stratagene |
| pEX100T | Ampr oriT sacB lacPOZ′ | 34 |
| pGEM-Teasy | Ampr lacPOZ′ | Promega |
| pPS856 | Ampr Gmr FRT | 17 |
| pEXpslA | pEX100T containing GM cassette flanked by 5′ and 3′ ends of the pslA gene | |
| pTZ110 | Broad-host-range lacZ fusion vector | 33 |
| pBBR1-MCS5 | Gmr lacPOZ′ mob, broad-host-range vector | 20 |
| pBBR1-MCS5::pslA | pBBR1-MCS5 containing EcoRI/BamHI PCR fragment of pslA | This study |
| pSKPpsl | pBluescript SK(−) containing EcoRI/BamHI PCR fragment of psl promoter | This study |
| pTZ110::Ppsl | pTZ110 containing EcoRI/BamHI PCR fragment of psl promoter | This study |
| pProbeAT′ | Broad-host-range gfp (stable form) fusion vector | 24 |
| pProbeAT′::Ppsl | pProbeAT′ containing EcoRI/BamHI PCR fragment of psl promoter | This study |
| pProbe′-gfp[AAV] | Broad-host-range gfp (unstable form) fusion vector | |
| pProbe′-gfp[AAV]::Ppsl | pProbe′-gfp[AAV] containing EcoRI/BamHI PCR fragment of psl promoter | 24 |
| pProbe′-gfp[AAV]::ΩGm-Ppsl | pProbe′-gfp[AAV]::Ppsl containing a gentamicin restistance gene cassette in the opposite direction to that of the gfp gene | This study |
| pGEM-Teasy::RACE | pGEM-Teasy containing 5′-RACE PCR product | This study |
| Oligonucleotides | ||
| PA2231-KO-UA | GTAGATAGCCTGTCGCTGACACGGG | This study |
| PA2231-KO-DA | AAAAAGGATCCGATCACCGTCCGTTGCAGGTACATG | This study |
| PA2231-KO-UB | AAAAAGGATCCCGGCTGATCTGGGTCTTCAAGTTCC | This study |
| PA2231-KO-DB | TCAGTAGACTTCCTTGGTCAGGAG | This study |
| PA2231-UE | AAAAAGAATTCTGGACTGCCCGTGATCGGCAGAGC | This study |
| PA2231-DB | AAAAAGGATCCGCAGGGGATCAGCGGGGCGACGGC | This study |
| PpslUE | AAAAAGAATTCCCGAAATGGCACGAGGCGGCGCTG | This study |
| PpslDB | AAAAAGGATCCGCCGATCACGGGCAGTCCATTGA | This study |
| RACE-primer1 | CAGGCCCAGGCCCTGGTACATGAACAACAGCAGGC | This study |
| RACE-primer2 | CGTCGGAGTAGACGTCCAGTGCCTGGAAC | This study |
| RACE-primer3 | GCGCAGCTCCACGGGAACCAGGCCGGGG | This study |