TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference(s) or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | F′ Φf80dlacZΔM15Δ(lacZYA-argF)recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 deoR U169 | Life Technologies |
| SM10 | Mobilizing strain; RP4 tra genes integrated in chromosome; Kmr | 40 |
| B. cepacia | ||
| K56-2 | Clinical isolate; genomovar III | 9, 23, 25 |
| K56orbA::tp | orbA::tpr derivative of K56-2 | This study |
| Plasmids | ||
| pNOT19 | Modified pUC19 cloning vector; Apr | 37 |
| pUCP26 | Broad-host-range vector; IncP OriT; pRO1600 ori; Tcr | 38 |
| pPD519 | pNOT19 with 6.1-kb SphI fragment from K56-2 containing the pvdA, orbA, and pvdF genes | 42 |
| pEX18Tc | Suicide vector; sacB Tcr | 20 |
| pEX18Ap | Suicide vector; sacB Apr | 20 |
| p34E-Tp | Source of tp cassette; Tpr | 12 |
| pRK2013 | ColE1 Tra (RK2)+ Kmr | 14 |
| pCC524 | pEX18TC with a 3-kb EcoRI-BglII fragment containing orbA cloned into the EcoRI-BamHI sites | This study |
| pCC524T | pCC524 with a SalI fragment containing the 0.6-kb Tpr cassette from p34E-Tp inserted in the XhoI site in the orbA gene | This study |
| pPD524 | pEX18Ap with a 3-kb EcoRI-BglII fragment containing orbA cloned into the EcoRI-BamHI sites | This study |
| pPD526 | pUCP26 with a 3-kb EcoRI-HindIII fragment from pPD524 cloned into the EcoRI-HindIII sites | This study |