TABLE 1.
Strains and plasmids used in this study
| Strain/plasmids | Relevant characteristicsa | Source/reference |
|---|---|---|
| Strains | ||
| B. cenocepacia | ||
| K56-2 | ET12 clone related to J2315, CF clinical isolate | BCRRCb (46) |
| K56-2 ΔatsR | Deletion of atsR | (34) |
| K56-2 ΔcepI | Deletion of cepI | (16) |
| K56-2 ΔatsRΔcepI | Deletion of cepI in K56-2 ΔatsR | (16) |
| K56-2 ΔatsR atsR+ | atsR integration in K56-2 ΔatsR | This study |
| K56-2 ΔatsRΔcepI atsR+ | atsR integration in K56-2 ΔatsRΔcepI | This study |
| K56-2 ΔatsRΔcepI atsRH245A+ | atsRH245A integration in K56-2 ΔatsRΔcepI | This study |
| K56-2 ΔatsRΔcepI atsRD536A+ | atsRD536A integration in K56-2ΔatsRΔcepI | This study |
| K56-2 ΔatsRΔcepI atsRΔRD+ | atsRΔRD integration in K56-2 ΔatsRΔcepI | This study |
| K56-2 ΔatsR atsRH245A+ | atsRH245A integration in K56-2 ΔatsR | This study |
| K56-2 ΔatsR atsRD536A+ | atsRD536A integration in K56-2ΔatsR | This study |
| K56-2 ΔatsR atsRΔRD+ | atsRΔRD integration in K56-2 ΔatsR | This study |
| K56-2 ΔatsRΔcepI +ΔatsT atsRD536A | Deletion of atsT in K56-2 ΔatsRΔcepI atsRD536A+ | This study |
| K56-2 ΔatsRΔcepI +ΔatsT atsRΔRD | Deletion of atsT in K56-2 ΔatsRΔcepI atsRΔRD+ | This study |
| K56-2 ΔatsR ΔatsT atsRD536A+ | Deletion of atsT in K56-2 ΔatsR atsRD536A+ | This study |
| K56-2 ΔatsR ΔatsT atsRΔRD+ | Deletion of atsT in K56-2 ΔatsR atsRΔRD+ | This study |
| K56-2 ΔcepI ΔatsT | Deletion of atsT in K56-2 ΔcepI | This study |
| K56-2 ΔcepI ΔatsT atsT + | atsT integration in K56-2 ΔcepI ΔatsT | This study |
| K56-2 ΔcepI ΔatsT atsTD208A+ | atsTD208A integration in K56-2 ΔcepI ΔatsT | This study |
| K56-2 ΔatsRΔhcp | Deletion of hcp in K56-2 ΔatsR | (34) |
| E. coli | ||
| DH5α | F− φ80lacZ M15 endA1 recA1 supE44 hsdR17 (rK− mK+) deoR thi-1 nupG supE44 gyrA96 relA1 Δ (lacZYA-argF)U169, λ− | Laboratory stock |
| GT115 | F− mcrA Δ(mrr-hsdRMS-mcrBC) φ80ΔlacZΔM15 ΔlacX74 recA1 rpsL (StrA) endA1Δdcm uidA(ΔMluI)::pir-116 ΔsbcC-sbcD | Invivogen |
| BL21 | F− dcm ompT lon hsdS (rB− mB−) gal λ (DE3) | Laboratory stock |
| Plasmids | ||
| pAtsRChr | atsR cloned in pMH447 for chromosomal complementation | This study |
| pAtsRH245AChr | atsRH245A cloned in pMH447 for chromosomal complementation | This study |
| pAtsRD536AChr | atsRD536A cloned in pMH447 for chromosomal complementation | This study |
| pAtsRΔRDChr | atsRΔRD cloned in pMH447 for chromosomal complementation | This study |
| pAtsTChr | atsT cloned in pMH447 for chromosomal complementation | This study |
| pAtsTD208AChr | atsTD208A cloned in pMH447 for chromosomal complementation | This study |
| pDAI-SceI-SacB | oripBBR1, TetR, Pdhfr, mob+, expressing I-SceI, SacB | (31) |
| pET28a(+) | Cloning vector, IPTG inducible for N-terminal His6 fusion, Kanr | Laboratory stock |
| pDelatsT | pGPI-SceI with fragments flanking atsT | This study |
| pDelM0378 | pGPI-SceI with fragments flanking bcam0378 | This study |
| pMK1 | pET28a (+) encoding B. cenocepacia bcam0378, His6: kanR | This study |
| pMK2 | pET28a (+) encoding B. cenocepacia atsT, His6: kanR | This study |
| pMK4 | pET28a (+) encoding B. cenocepacia atsR-RDD536A, His6: kanR | This study |
| pMK5 | pET28a (+) encoding B. cenocepacia atsRD536A, His6: kanR | This study |
| pMZ24 | pET28a (+) encoding B. cenocepacia atsR205–605, His6: kanR | M. Al-Zayer |
| pMZ25 | pET28a (+) encoding B. cenocepacia atsR, His6: kanR | M. Al-Zayer |
| pMZ33 | pET28a (+) encoding B. cenocepacia atsR-HK, His6: kanR | M. Al-Zayer |
| pMZ34 | pET28a (+) encoding B. cenocepacia atsR-RD, His6: kanR | M. Al-Zayer |
| pMZ36 | pET28a (+) encoding B. cenocepacia atsR atsRH245A, His6: kanR | M. Al-Zayer |
| pMH447 | pGPI-SceI derivative used for chromosomal complementation | (30) |
| pRK2013 | oricolE1, RK2 derivative, KanR, mob+, tra+ | (28) |
a kanR, kanamycin resistance; TetR, tetracycline resistance.
b B. cepacia Research and Referral Repository for Canadian CF Clinics.