TABLE 1.
Bacterial strains, plasmids, and transposons
| Strain, plasmid, or transposon | Genotype or descriptiona | Source or reference |
|---|---|---|
| B. cenocepacia strains | ||
| 715j | CF isolate, prototroph | 15 |
| KLF1 | Spontaneous Pch− mutant of 715j | 21 |
| OM1 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | This study |
| OM3 | KLF1-orbS::mini-Tn5Tp (Orb− Pch−) | This study |
| OM4 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | This study |
| OM6 | KLF1-orbJ::mini-Tn5Tp (Orb− Pch−) | This study |
| OM11 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | This study |
| OM12 | KLF1-orbJ::mini-Tn5Tp (Orb− Pch−) | This study |
| OM13 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | This study |
| OM15 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | This study |
| OM17 | KLF1-orbI::mini-Tn5Tp (Orb− Pch−) | |
| 715j-orbS::Tp | 715j with dfr cassette inserted in orbS (Orb− Pch+) | This study |
| E. coli strains | ||
| MC1061 | hsdR araD139 Δ(ara-leu)7697 ΔlacX74 galU galK rpsL (Smr) | 9 |
| JM83 | F−ara Δ(lac-proAB) rpsL φ80dlacZΔM15 (Smr) | 89 |
| H1717 | aroB fhuF::λplacMu53 | 25 |
| QC1732 | F− Δ(argF-lac)U169 rpsL Δfur::kan (Smr Kmr) | 76 |
| CC118(λpir) | araD139 Δ(ara-leu)7697 ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(am)recA1 λpir (Rfr) | 28 |
| S17-1 | thi proA hsdR recA RP4-2-tet::Mu-1 kan::Tn7 integrant (Tpr Smr) | 67 |
| S17-1(λpir) | S17-1 lysogenized with λpir | 67 |
| BW19851 | ΔuidA::pir+recA1 hsdR17 creB510 endA1 zbf-5 thi RP4-2-tet::Mu-1 kan::Tn7 integrant (Tpr Smr) | 49 |
| Plasmids and transposons | ||
| pBBR1MCS | Mobilizable BHR cloning vector; IncP and ColE1 compatible (Cmr) | 35 |
| pBBR1MCS-orbS | pBBR1MCS containing the B. cenocepacia 715j orbS gene (Cmr) | This study |
| pBBR1MCS-orbS::Tp | pBBR1MCS-orbS with the dfr (Tpr) cassette inserted in orbS (Cmr) | This study |
| pPR9TT | Mobilizable BHR lacZ translational fusion vector, IncP (Cmr Apr) | 62 |
| pUC18NotlacZYA | pUC18Not containing 5.89-kb promoterless lacZYA cassette (Apr) | E. Dave and M. S. Thomas, unpublished data |
| pKAGd4 | Transcriptional fusion vector derived from pPR9TT (Cmr Apr) | This study |
| pKAGd4-porbS | pKAGd4 containing porbS-lacZ fusion (Cmr Apr) | This study |
| pKAGd4-porbH | pKAGd4 containing porbH-lacZ fusion (Cmr Apr) | This study |
| pKAGd4-porbE | pKAGd4 containing porbE-lacZ fusion (Cmr Apr) | This study |
| pKAGd4-porbI | pKAGd4 containing porbI-lacZ fusion (Cmr Apr) | This study |
| pKAGd4ΔAp | pKAGd4 containing nptII cassette from p34E-Km inserted in bla gene (Cmr Kmr) | This study |
| pKAGd4ΔAp-porbS | pKAGd4ΔAp containing porbS-lacZ fusion (Cmr Kmr) | This study |
| pKNOCK-Km | Mobilizable suicide vector containing nptII cassette (Kmr) | 2 |
| p34E-Tp | p34E containing dfr gene (Tpr Apr) | 19 |
| p34E-Km | p34E containing nptII cassette from pKNOCK-Km (Kmr Apr) | E. H. Bull and M. S. Thomas, unpublished data |
| pTrc99A | pBR322-derived expression vector containing strong inducible trc promoter and lacIq gene (Apr) | 4 |
| pAHA21 | pTrc99A containing B. cenocepacia fur gene (Apr) | 42 |
| pUC18 | E. coli-specific cloning vector (Apr) | 89 |
| pBluescript II KS | E. coli-specific cloning vector (Apr) | 3 |
| p3ZFBS | pGEM3Z containing consensus E. coli Fur-binding site (Apr) | 81 |
| pBS-porbS | pBluescript KS II containing full-length orbS promoter (Apr) | This study |
| pBS-porbH-69 | pBluescript KS II containing −69 orbS promoter (Apr) | This study |
| pBS-porbS-40 | pBluescript KS II containing −40 orbS promoter (Apr) | This study |
| pBS-porbS+5 | pBluescript KS II containing +5 orbS promoter (Apr) | This study |
| pBS-porbH | pBluescript KS II containing orbH promoter (Apr) | This study |
| pBS-porbE | pBluescript KS II containing orbE promoter (Apr) | This study |
| pBS-porbI | pBluescript KS II containing orbI promoter (Apr) | This study |
| pUTmini-Tn5Cm | Mobilizable suicide vector, pUT, containing mini-Tn5Cm (Cmr Apr) | 17, 18 |
| pUTmini-Tn5Tp | Ω-Cm interposon of pUTmini-Tn5Cm replaced by dfr cassette of p34E-Tp (Tpr Apr) | This study |
| pSHAFT | pUTmini-Tn5Cm containing deletion of BglII fragment harboring tnp gene and I end of mini-Tn5Cm (Cmr Apr) | 66 |
| pSHAFT-orbS::Tp | pSHAFT containing the SalI-NotI orbS::Tp fragment from pBBR1MCS-orbS::Tp (Cmr Apr Tpr) | This study |
a
Apr, ampicillin resistance; Cmr, chloramphenicol resistance; Kmr, kanamycin resistance; Rfr, rifampicin resistance; Smr, streptomycin resistance; Tpr, trimethoprim resistance; Pch−, pyochelin-negative phenotype; Orb−, ornibactin-negative phenotype; BHR, broad host range.