TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference(s) |
|---|---|---|
| A. tumefaciens A136 | Ti plasmidless host | C. Fuqua |
| E. coli strains | ||
| DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF) recA1 endA gyrA96 thi-1 hsdR17 supE44 relAl deoR(U169) | Invitrogen |
| TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL(Str) endA1 nupG | Invitrogen |
| HB101 | supE44 hsdS20 (rB mB) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 79 |
| MM294 | F−endA1 hsdR17 supE44(AS) rfbD1 spoT1 thi-1 | 66 |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10(Tcr)] | Stratagene |
| Bcc strains | ||
| B. cepacia ATTC 25416T | Onion isolate | 60 |
| B. multivorans strains | ||
| LMG13010T | CF isolate | 60 |
| C5393 | CF isolate | 60 |
| B. cenocepacia strains | ||
| J2315 | CF isolate, BCESM+cblA+ | 60 |
| BC7 | CF isolate, BCESM+cblA+ | 60 |
| C5424 | CF isolate, BCESM+cblA+ | 60 |
| C6433 | CF isolate, BCESM+ | 60 |
| C1394 | CF isolate BCESM+ | 60 |
| PC184 | CF isolate, BCESM+ | 60 |
| CEP511 | CF isolate, BCESM+ | 60 |
| J415 | CF isolate, BCESM− | 60 |
| ATTC 17765 | Urinary tract infection isolate, BCESM+ | 60 |
| PC715j | CF isolate, BCESM− | 65 |
| H111 | CF isolate, BCESM− | 75 |
| H111-R | cepR::Km derivative of H111 | 35 |
| H111-R2 | cepR2::pEX18Gm derivative of H111 | This study |
| H111-R/R2 | Kmr Gmr, cepR2::pEX18Gm mutant of H111-R | This study |
| K56-2 | CF isolate, BCESM+cblA+ | 60 |
| K56-2cepR2a | cepR2::Tp derivative of K56-2, clone a, Tpr | This study |
| K56-2cepR2b | cepR2::Tp derivative of K56-2, clone b, Tpr | This study |
| K56-R2 | cepR::Tn5-OT182 derivative of K56-2, Tcr | 52 |
| K56-dI2 | ΔcepI derivative of K56-2 | 63 |
| K56-2cciI | cciI::Tp derivative of K56-2, Tpr | 5 |
| K56-2cciR | cciR::Tp derivative of K56-2, Tpr | 63 |
| K56-2cciIR | ΔcciIR derivative of K56-2 | 63 |
| K56-2cepRcciIR | cepR::Tp, ΔcciIR derivative of K56-2, Tpr | 63 |
| B. stabilis strains | ||
| LMG14294 | CF isolate | 60 |
| LMG14086 | Respiratory isolate | 60 |
| B. vietnamiensis strains | ||
| PC259 | CF isolate | 47 |
| G4 | Water treatment facility isolate | 71 |
| B. dolosa strains | ||
| LMG19468T | CF isolate | J. LiPuma |
| LMG18943 | CF isolate | 8 |
| B. ambifaria strains | ||
| CEP0996 | CF isolate | 9 |
| LMG17828 | Soil isolate | 9 |
| B. athina strains | ||
| LMG20980T | Soil isolate | 10 |
| LMG20982 | Hospital environmental isolate | D. P. Speert |
| B. pyrrocinia LMG 21822 | Soil isolate | 10 |
| Plasmids | ||
| pEX18Tc | Gene replacement vector, oriT+sacB+ Tcr | 32 |
| pUCP26 | Broad-host-range vector, Tcr | 99 |
| pCR2.1Topo | Cloning vector for PCR products, Apr Kmr | Invitrogen |
| pRK2013 | ColEl Tra (RK2)+, Kmr | 21 |
| pRK415 | Broad-host-range vector, Tcr | 39 |
| p34E-Tp | Source of Tp resistance cassette, Tpr | 13 |
| p34S-Tc | Source of Tc resistance cassette, Tcr | 12 |
| pCF218 | IncP plasmid expressing TraR, Tcr | 100 |
| pCF372 | pUCD2 with a traI-lacZ fusion, Spr | 25 |
| pRM1T6 | pCR2.1TOPO containing the 1.6-kb cepR2 fragment, Apr Kmr | This study |
| pRM613 | pUCP26 containing a 1.3-kb SstI/SmaI fragment from pRM1T6 including cepR2, Tcr | This study |
| pRM516 | pRK415 containing the 1.6-kb cepR2 fragment from pRM1T6, Tcr | This study |
| pBS13 | zmpA::luxCDABE transcriptional fusion constructed in pMS402, Kmr Tpr Tcr | B. Subsin |
| pBS9 | zmpB::luxCDABE transcriptional fusion constructed in pMS402, Kmr Tpr Tcr | 42 |
| pBBR1MCS | Cmr, broad-host-range cloning vector | 44, 45 |
| pBBR-cepR | Cmr, pBBR1MCS containing the cepR gene of B. cenocepacia H111 | S. Schmidt |
| pEX18Gm | Gene replacement vector, oriT+sacB+, Gmr | 32 |
| pJBA89 | Apr, pUC18Not-luxR-PluxI-RBSII-gfp(ASV)-T0-T1 | 3 |
| pJBA89luxR− | Apr, pUC18Not-PluxI-RBSII-gfp(ASV)-T0-T1 | This study |
| pJTR2 | Cmr, pBBR1MCS containing the cepR2 gene of B. cenocepacia H111 | This study |
| pRK600 | Cmr, ColE1 oriV RK2-Mob+ RK2-Tra+ | 40 |
| pRK2013 | Kmr, RK2 derivative, Mob+ Tra+ ColE1 | 15 |
| pSU11 | Gmr, broad-host-range lacZ-based promoter probe vector | L. Eberl |
| pRN3 | Tpr, broad-host-range lacZ-based promoter probe vector, derivative of pSU11 | This study |
a
BCESM, B. cepacia epidemic strain marker; cblA, cable pilus gene; Ap, ampicillin; Cm, chloramphenicol; Km, kanamycin; Sp, spectinomycin; Tp, trimethoprim; Tc, tetracycline.