TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| B. cepacia | ||
| BC11 | Wild-type B. cepacia soil isolate | 16 |
| BC11R | Spontaneous Rifr mutant of BC11 | This work |
| AFC143, AFC145, AFC202, AFC666 | afc::Tn5 derivatives of BC11R Rifr Kmr | This work |
| AFC666M2 | afcA::nptI derivative of BC11R made by allelic exchange | This work |
| E. coli | ||
| DH5α | endA1 hsdR trecA1 gyrA relA φ80lacZΔM15 | 42 |
| SM10 | RP4-2-Tc::Mu recA thi pro his | 58 |
| HB101 | recA rpsL ara galK leu hsdS Δ(gpt-proA) supE xyl | 42 |
| Plasmids | ||
| pTZ19U | ColE1 replicon, Apr | 45 |
| pRK415 | IncP1 replicon, Mob+ Tcr | 29 |
| pTOK2 | ColE1 replicon, suicide plasmid, Tcr | 33 |
| pSUP2021 | ColE1 replicon, suicide plasmid with Tn5, Kmr | 58 |
| pSB315 | pUC4K derivative with nonpolar nptI cartridge, Kmr Apr | 14 |
| pRK2013 | ColE1 replicon, Tra+-mobilizing plasmid, Kmr | 9 |
| pKW666 | 10-kb EcoRI genomic fragment with afc::Tn5 of AFC666 cloned into the EcoRI site of pTZ19U, Kmr Apr | This work |
| pT666 | 10-kb EcoRI fragment of pKW666 cloned into the EcoRI site of pTOK2, Tcr Kmr | This work |
| pKW666-1 | 5.6-kb BamHI-EcoRI fragment of pKW666 cloned in the BamHI and EcoRI sites of pTZ19U, Kmr Apr | This work |
| pKW666-2 | 2.8-kb PstI-EcoRI fragment of pKW666-1 cloned in the PstI and EcoRI sites of pTZ19U, Apr | This work |
| pKW666-4 | 3.0-kb HpaI-EcoRI fragment of pKW666-1 cloned in pTZ19U, Apr | This work |
| pKW666-5 | 0.9-kb SalI fragment with nonpolar nptI from pSB315 cloned into the XhoI site of pKW666-4, Kmr Apr | This work |
| pT666-1 | 3.9-kb XbaI-EcoRI fragment of pKW666-5 cloned into the XbaI and EcoRI sites of pTOK2, Tcr Kmr | This work |
| pR666 | 2.2-kb StyI-EcoRI fragment of pKW666-1 cloned into pRK415, Tcr | This work |
a
Tcr, Kmr, Rifr, and Apr denote resistance to tetracycline, kanamycin, rifampin, and ampicillin, respectively.