TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype, phenotype, or descriptiona | Reference or source |
|---|---|---|
| Strains | ||
| Acinetobacter | ||
| ADP1 (BD413) | Wild type | 22 |
| ACN73 | areR::Kmr; transformation of ADP1 with pBAC87 | This study |
| ADPW56 | areC::Kmr; transformation of ADP1 with pADPW22 | This study |
| ADPW57 | areB::Kmr; transformation of ADP1 with pADPW26 | This study |
| ADPW58 | areA::Kmr; transformation of ADP1 with pADPW37 | This study |
| ADPW68 | areB::Kmr; transformation of ADP1 with pADPW75 | This study |
| ISA36 | benM::Smr Spr | 9 |
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17 (rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | Gibco BRL |
| XL1-Blue MRF′ | Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr)] | Stratagene |
| BL21(DE3)pLysS | F−ompT hsdSB(rB− mB−) dcm gal (DE3) pLysS Cmr | Promega |
| Plasmids | ||
| pET5a | Apr; T7 expression vector | Promega |
| pUC18 and pUC19 | Apr; cloning vector | 40 |
| pUC4K | Apr Kmr; source plasmid for Kmr cassette | 36 |
| pUI1637 | Apr Kmr; source plasmid for Kmr cassette | 14 |
| pBAC68 | 7.0-kbp BglII/NsiI fragment cloned from ISA36 containing Smr Spr cassette and all of benP and part of areR in pUC19 | 8 and this study |
| pBAC78 | 0.86-kbp BglII/KpnI fragment containing part of areR in pUC19 | This study |
| pBAC87 | pBAC78 with Kmr cassette excised from pUI1637 with EcoRV and cloned into SnaBI site in areR | This study |
| pBAC98 | 6.7-kbp NsiI/KpnI fragment cloned from ACN73 containing Kmr cassette, all of areCB, and part of areR and areA | This study |
| pADPW21 | 1.6-kbp BamHI*/SacI fragment containing areC in pUC18 | This study |
| pADPW22 | pADPW21 with Kmr cassette from pUC4K cloned into BglII site in areC | This study |
| pADPW25 | 1.4-kbp BamHI*/EcoRI* fragment containing areB in pUC18 | This study |
| pADPW26 | pADPW25 with Kmr cassette from pUC4K cloned into BclI site in areB | This study |
| pADPW29 | NdeI*/EcoRI* fragment containing areC in pET5a | This study |
| pADPW30 | NdeI*/EcoRI* fragment containing areB in pET5a | This study |
| pADPW31 | 0.65-kbp SacI*/SalI* fragment containing part of areA in pUC18 | This study |
| pADPW32 | 8.0-kbp SacI fragment cloned from ADPW57 containing Kmr cassette and all of areA in pUC18 | This study |
| pADPW33 | 3.6-kbp HindIII subclone of pADPW32 in pUC18 | This study |
| pADPW36 | 1.2-kbp EcoRI* fragment containing areA in pUC18 | This study |
| pADPW37 | pADPW36 with Kmr cassette from pUC4K cloned into NsiI site in areA | This study |
| pADPW40 | NdeI*/EcoRI* fragment containing areA in pET5a | This study |
| pADPW72 | 1.0-kbp EcoRI*/HindIII* fragment containing part of areB in pUC18 | This study |
| pADPW75 | pADPW72 with Kmr cassette from pUI1637 cloned into SacI site in areB | This study |
a
Restriction sites added by PCR are indicated by superscript asterisks.