TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype and/or phenotype | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | supE44 lacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Bethesda Research Laboratories |
| S17-1 | Integrated RP4-2, Tc::Mu Km::Tn7 | 44 |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac (F′ proAB lac1qZΔM15 Tn10) | Stratagene |
| B. cepacia strains | ||
| J2315 | CF clinical isolate, genomovar III | 17 |
| D9 | fliG::mini-Tn5Tc derivative of J2315 | This study |
| CM58 | fliI::cat derivative of J2315 | This study |
| CM100 | CM58 with plasmid pJG7 | This study |
| CM340 | D9 with plasmid pMT46 | This study |
| Plasmids | ||
| pBluescript SK(−) | Cloning and single-stranded phagemid | Stratagene |
| pBluescript SK(+) | Cloning and single-stranded phagemid | Stratagene |
| pGEM-T Easy | TA cloning vector | Promega |
| pLAFR-5 | Cosmid vector, Tcr | 27 |
| pCAT1 | Source of cat cassette, Cmr | This study |
| pMR4 | Broad-host-range vector, Tcr | C. Mohr and R. Roberts |
| pUCP18 | Broad-host-range vector, Apr | 52 |
| pCMT1 | Cmr derivative of pUCP18 | This study |
| pCM117 | pLAFR-5 cosmid containing 20-kb B. cepacia genomic DNA insert encoding the fliG locus | This study |
| pAR1 | 3.5-kb XhoI fragment from pCM117 subcloned into pBluescript SK(+) | This study |
| pAR2 | 1.3-kb PstI fragment containing fliI, subcloned from pAR1 into pBluescript SK(+) | This study |
| pAR3 | 1.3-kb XhoI-PstI fragment from pAR2 subcloned into pBluescript SK(+) | This study |
| pAR4 | pAR3 containing fliI gene inactivated with cat cassette, Cmr/Apr | This study |
| pJG7 | 1.6-kb PstI fragment encoding fliI truncated at codon 476, cloned in pMR4 | This study |
| pMT38 | 1.5-kb PCR product encoding fliG cloned into pGEM-T Easy | This study |
| pMT46 | 1.5-kb fliG PCR product cloned into pCMT1 | This study |