TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Source |
|---|---|---|
| Strains | ||
| E. coli | ||
| TG1 | supE Δ(lac-proAB) thi hsdRΔ5 (F′ traD36 rpoA+B+lacIqZΔM15) | Lab collection |
| TOP10F′ | F′ [lacIq Tn10 (Tetr)] mrcA Δ(mrr-hsdRMS-mcrBC) φ80 lacZΔM15 ΔlacX74 recA1 | Invitrogen |
| S17-1 | Conjugative helper strain; galU galK rpsL (Strr) endA1 nupG thi pro hsdR hsdM recA (RP4-2Tc::Mu Km::Tn7)λpir | Lab collection |
| CC118(λpir) | Host strain for pKNG101 replication; Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 Rfr (λpir) | Lab collection |
| P. aeruginosa | ||
| PAO1 | Wild type | Lab collection |
| PAO1Δflp | PAO1 deletion mutant for flp gene | This study |
| PAO1ΔtadA | PAO1 deletion mutant for tadA gene | This study |
| PAO1ΔrcpA | PAO1 deletion mutant for rcpA gene | This study |
| PAO1ΔfppA | PAO1 deletion mutant for fppA gene | This study |
| PAO1ΔpilD | PAO1 deletion mutant for pilD gene | Lab collection |
| PAO1ΔfliCΔpilA | PAO1 deletion mutant for fliC and pilA genes | Lab collection |
| Plasmids | ||
| pCR2.1 | TA cloning vector for PCR products; lacZα ColE1 f1 ori Apr Kmr | Invitrogen |
| pMMB190 | Broad-host-range vector; IncQ ptac lacZα Apr | Lab collection |
| pBBR1MCS-2 | Broad-host-range vector; Kmr | Lab collection |
| pKNG101 | Suicide vector in P. aeruginosa; sacB Strr | Lab collection |
| pMMBflp | flp gene cloned in pMMB190 at BamHI/HindIII | This study |
| pBBRfppA | fppA gene cloned in pBBR1MCS-2 at EcoRI | This study |
| pBBRfppAD17A | fppA gene with D17A substitution cloned into pBBR1MCS-2 | This study |
| pBBRfppAD78A | fppA gene with D78A substitution cloned into pBBR1MCS-2 | This study |
| pBBRfppAD17AD78A | fppA gene with D17A and D78A substitutions cloned into pBBR1MCS-2 | This study |
| pDEST14 | Destination vector of the Gateway system, T7 terminator, T7 promoter; Cmr Apr | Invitrogen |
| pLAFR3 | Cosmid, IncP, lac promoter; Tcr | Lab collection |
| pLAFRrcpA | rcpA gene cloned into pLAFR3 at BamHI/HindIII | This study |
| pLAFRtadA | tadA gene cloned into pLAFR3 at BamHI/HindIII | This study |
| pPRK2013 | ColE1 ori tra+mob+ Kmr | Lab collection |
a
Strr, streptomycin resistance; Apr, ampicillin resistance; Kmr, kanamycin resistance; Tcr, tetracycline resistance.