TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference |
|---|---|---|
| Strains | ||
| A. tumefaciens | ||
| A136 | Ti plasmidless host | C. Fuqua |
| E. coli | ||
| TOP10 | F−mcrAΔ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Str) endA1 nupG | Invitrogen |
| DH10B | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara-leu)7697 galU galK λ−rpsL nupG | Invitrogen |
| SY327 | araD Δ(lac-pro) argE(Am) recA56 RifrnalA λpir | 39 |
| B. cenocepacia | ||
| K56-2 | CF isolate | 34 |
| K56-R2 | cepR::Tn5-OT182 derivative of K56-2, Tcr | 29 |
| K56-2ΔcepR | cepR derivative of K56-2 | 41 |
| K56-2shvR::Tp | shvR::Tp derivative of K56-2, Tpr | This study |
| K56-2ΔshvR | shvR derivative of K56-2 | This study |
| K56-2cepIcciIb | ΔcepI cciI::Tp derivative of K56-2, Tpr | 36 |
| Plasmids | ||
| pCR2.1Topo | Cloning vector for PCR products, Apr Kmr | Invitrogen |
| pCF218 | IncP plasmid expressing TraR, Tcr | 63 |
| pMV26 | traI-luxCDABE fusion, Kmr | 51 |
| pRK2013 | ColEl Tra (RK2)+, Kmr | 15 |
| pGSVTplux | Mobilizable suicide vector containing lux operon, derivative from pGSV3-lux, OriT, Tpr | 4 |
| pEX18Tc | Gene replacement vector oriT+, sacB+, Tcr | 20 |
| pUCP26-BCAS0225 | pUCP26 with 1.7-kb PstI-BamHI fragment containing BCAS0225 and upstream region, Tcr | 4 |
| pEX18Tc-BCAS0225::Tp | pEX18Tc containing BCA0225 inactivated with a Tp cassette from pGSVTplux | This study |
| pGPI-SceI | oriR6K, Tpr, mob+, carries I-SceI endonuclease recognition site, Tpr | 16 |
| pDAI-SceI | pDA17 carrying the I-SceI gene, Tcr | 16 |
| pUCP28T | Broad-host-range vector, Tpr | 48 |
| p28T-shvR | pUCP28T with 1.7-kb PstI-BamHI fragment containing BCAS0225 and upstream region, Tcr | This study |
| pSLR100 | pUCP28T with 1.65-kb KpnI-SphI fragment from pSLA3.2 containing cepR gene, Tpr | 29 |
| pMS402 | lux-based promoter reporter plasmid, Kmr Tpr | 13 |
| pCP300 | cepI::lux transcriptional fusion constructed in pMS402, Kmr Tpr | 36 |
| pRM432 | cepR::lux transcriptional fusion constructed in pMS402, Kmr Tpr | 36 |
| pRM445 | cciIR::lux transcriptional fusion constructed in pMS402, Kmr Tpr | 36 |
| pPafcA | afcA::lux transcriptional fusion constructed in pMS402, Kmr Tpr | 41 |
| pPafcC | afcC::lux transcriptional fusion constructed in pMS402, Kmr Tpr | This study |
| pBS13 | zmpA::lux transcriptional fusion constructed in pMS402, Kmr Tpr Tcr | 37 |
| pBS9 | zmpB::lux transcriptional fusion constructed in pMS402, Kmr Tpr Tcr | 26 |
| pPgspC | gspC::lux transcriptional fusion constructed in pMS402, Kmr Tpr | This study |
| pPgspG | gspG::lux transcriptional fusion constructed in pMS402, Kmr Tpr | This study |
| pPshvR | shvR::lux transcriptional fusion constructed in pMS402, Kmr Tpr | This study |
| pCS26-Pac | lux-based promoter reporter plasmid, Kmr | 6 |
| pEPO100 | pCS26-Pac with 269-bp XhoI-BamHI fragment from pCP300 containing cepI promoter, Kmr | This study |
| pEPO101 | pCS26-Pac with 371-bp XhoI-BamHI fragment from pPafcA containing afcA promoter, Kmr | This study |
| pEPO128 | pCS26-Pac with 405-bp XhoI-BamHI fragment from pPafcC containing afcC promoter, Kmr | This study |