TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| TG1 | F′ [traD36 proAB+ lacIq lacZΔM15] Δ(lac-proAB) | Laboratory collection |
| CC118λpir | (ara-leu) araD lacX74 rpsE rpoB argE(Am) recA1 RfR(λpir) | 28 |
| DH10B | F− endA1 recA1 nupG rpsL ΔlacX74 ϕ80lacZΔM15 araD139Δ(ara leu)7697 mcrA Δ(mrr-hsdRMS-mcrBC) λ− | Invitrogen |
| TOP10F′ | Similar to DH10B, with the F plasmid containing lacIq and Tn10 | Invitrogen |
| BL21(DE3) | F− ompT gal lon λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | 62 |
| BTH101 | F′ cya araD galE galK hsdR2 mcrA1 mcrB1 relA1; Smr | 35 |
| P. aeruginosa | ||
| PA7 | Nonrespiratory clinical isolate | 26 |
| PA7 Δtxc | txcPQRSTUVWXYZ deletion mutant in PA7 | This work |
| PA7::lacZ | PA7 strain carrying the lacZ promoterless fusion inserted into the att site | This work |
| PA7::PcbpE-lacZ | PA7 strain carrying the PcbpE-lacZ fusion inserted into the att site | This work |
| PA7::Ptxc-lacZ | PA7 strain carrying the Ptxc-lacZ fusions inserted into the att site | This work |
| S. cerevisiae CRY1-2 | Cycloheximide resistant (cyh2R) and uracil prototrophic (ura3) | 33 |
| Plasmids | ||
| pCR2.1 | TA cloning; lacZ ColE1 f1 ori; Apr Kmr | Invitrogen |
| pRK2013 | Tra+ Mob+ ColE1; Kmr | 63 |
| pBBR1mcs4 | Cloning vector; Apr Cbr | 64 |
| pCbpE | pBBR1MCS-4 containing the cbpE-His6 DNA sequence; Apr Cbr | This work |
| pTtsSc | pBBR1MCS-4 containing the ttsSc-His6 DNA sequence; Apr Cbr | This work |
| pTtsRc | pBBR1MCS-4 containing the ttsRc-His6 DNA sequence; Apr Cbr | This work |
| pT7.5 | Cloning vector with the T7 promoter; Apr | 65 |
| pCbpD | pT7.5 containing the C-terminal cbpD-His10 DNA sequence; Apr | This work |
| pLLX13 | Yeast capture plasmid; URA3 CEN6 ARSH4 oriV incP tra oriT; Tcr | 33 |
| pLLX8 | Plasmid containing cyh2 and bla genes; cyh2R Apr Cbr | 33 |
| pCV1407-1420 | Plasmid containing TS1 and TS2 sequences and cyh2R and bla genes | This work |
| pRGP69 | pLLX13 containing the ttsS-txcZ DNA region | This work |
| pKNG101 | Suicide vector; oriR6K mobRK2 sacBR+; Smr | 66 |
| pHP45Ω-Tc | Plasmid containing the Tc resistance cassette; Apr Tcr | 67 |
| pKNG208 | Tcr suicide vector; oriR6K mobRK2 sacBR+; Tcr | This work |
| pKNG208Δtxc | pKNG208 containing the 500-bp upstream and the 500-bp downstream DNA regions of the txc cluster; Tcr | This work |
| MiniCTX-lacZ | Ω-FRT-attP-MCS ori int oriT; Tcr | 30 |
| pPtxc-lacZ | Mini-CTX containing the transcriptional fusion Ptxc-lacZ | This work |
| pPcbpE-lacZ | Mini-CTX containing the transcriptional fusion PcbpE-lacZ | This work |
| pUT18C | Expression vector encoding the T18 fragment of cyaA; Apr | 37 |
| pKT25 | Expression vector encoding the T25 fragment of cyaA; Cmr | 37 |
| pT18-TtsSc | pUT18C containing the TtsSc DNA sequence; Apr | This study |
| pT18-Pal | pUT18C containing the Pal DNA sequence; Apr | E. Bouveret |
| pT25-TtsR | pKT25 containing the TtsR DNA sequence; Cmr | This study |
| pT25-TolB | pKT25 containing the TolB DNA sequence; Cmr | E. Bouveret |
a
Smr, streptomycin resistance; cyh2R, cycloheximide resistance; Apr, ampicillin resistance; Kmr, kanamycin resistance; Cbr, carbenicillin resistance; Tcr, tetracycline resistance; Cmr, chloramphenicol resistance.