TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| Escherichia coli strains | ||
| DH5α | F− φ80 lacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17 (rK− mK+) supE44 thi-1 ΔgyrA96 relA1 | Laboratory stock |
| SY327 | araD Δ(lac pro) argE(Am) recA56 rifR nalA λ pir | 38 |
| SØ874 | lacZ trp Δ(sbcB-rfb) upp relA rpsL λ− | 40 |
| FAM3 | SØ874, ΔhldE1 Kmr | 37 |
| SAL7 | SØ874, ΔhldD Kmr | This study |
| Burkholderia cenocepacia strains | ||
| K56-2 | ET12 clone, CF clinical isolate | BCRRCb |
| SAL1 | K56-2, hldA::pSL5 Tpr | This study |
| SAL2 | K56-2, cysteine synthase B::pSL12 Tpr | This study |
| RSF19 | K56-2, wbxE::pRF201 Tpr | This study |
| Plasmids | ||
| pGP-ΩTp | oriR6K Tpr | This study |
| pRF200 | oriR6K Tpr | This study |
| pRF201 | pRF200, 320-bp wbxE mutagenesis fragment | This study |
| pRK2013 | RK2 derivative, Kmrmob+tra+oricolE1 | 17 |
| pSL5 | pGP-ΩTp, 273-bp hldA mutagenesis fragment | This study |
| pSL12 | pGP-ΩTp, 315-bp cysteine synthase B mutagenesis fragment | This study |
| pSCrhaB2 | pMLBAD with PRha (rhamnose-inducible promoter), Tpr | 9 |
| pSL3 | pSCrhaB2, B. cenocepacia hldA | This study |
| pSL6 | pSCrhaB2, chloramphenicol resistance cassette, Tpr Cmr | This study |
| pSL7 | pSL6, B. cenocepacia hldDA | This study |
| pSL8 | pSL6, B. cenocepacia hldA | This study |
| pSL9 | pSL6, B. cenocepacia hldD | This study |
a
Tp, trimethoprim; Cm, chloramphenicol; Km, kanamycin; Ap, ampicillin.
b
BCRRC, B. cepacia Complex Research and Referral Repository for Canadian CF Clinics.