TABLE 1.
Plasmids used in this studya
| Plasmid | Relevant characteristic | Origin |
|---|---|---|
| General | ||
| pUC118 | Vector: Apr; multiple cloning site in lacZ | Takara Shuzo Co. |
| pUC119 | Vector: Apr; multiple cloning site in lacZ | Takara Shuzo Co. |
| pTrc99A | Expression vector: Apr; multiple cloning site downstream of trc promoter | Amersham Pharmacia Biotech |
| pTrc6His | pTrc99A-derived expression vector: Apr; C-terminal 6-His tag | This study |
| Plasmids carrying MF-type transporter ORFs | ||
| pUCyajR | 2.0-kb KpnI-HindIII fragment containing yajR (putative transporter) gene cloned into pUC118, Apr | This study |
| pUCfsr | 1.8-kb EcoRI-SphI fragment containing fsr (fosmidomycin transporter) gene cloned into pUC118, Apr | This study |
| pUCmdfA | 1.6-kb KpnI-HindIII fragment containing mdfA (multidrug transporter) gene cloned into pUC118, Apr | This study |
| pUCyceE | 1.4-kb SphI-BamHI fragment containing yceE (putative transporter) gene cloned into pUC119, Apr | This study |
| pUCyceL | 1.7-kb SphI-BamHI fragment containing yceL (putative transporter) gene cloned into pUC119, Apr | This study |
| pUCydeA | 1.4-kb SphI-BamHI fragment containing ydeA (l-arabinose and IPTG transporter) gene cloned into pUC119, Apr | This study |
| pUCydeF | 1.7-kb SphI-BamHI fragment containing ydeF (putative transporter) gene cloned into pUC119, Apr | This study |
| pUCynfM | 2.5-kb SphI-BamHI fragment containing ynfL (putative regulator) and -M (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCydhC | 2.4-kb SphI-BamHI fragment containing ydhB (putative regulator) and -C (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCydiM | 1.7-kb SphI-BamHI fragment containing ydiM (putative transporter) gene cloned into pUC119, Apr | This study |
| pUCyebQ | 2.8-kb BamHI-EcoRI fragment containing kdgR (putative regulator) and yebQ (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCbcr | 2.3-kb AccI-KpnI fragment containing yeiD (putative regulator) and bcr (bicyclomycin transporter) genes cloned into pUC119, Apr | This study |
| pUCemrKY | 7.5-kb SphI-BamHI fragment containing evgS/A (two-component system) and emrK/Y (putative MFP/putative transporter) genes cloned into pUC119, Apr | This study |
| pUCemrAB | 3.9-kb SalI-BamHI fragment containing emrR (regulator), -A (MFP), and -B (multidrug transporter) genes cloned into pUC119, Apr | This study |
| pUCemrD | 2.0-kb HindIII-SacI fragment containing emrD (multidrug transporter) gene cloned into pUC119, Apr | This study |
| pUCyidY | 2.7-kb PstI-BamHI fragment containing yidY (putative transporter) and -Z (putative regulator) genes cloned into pUC119, Apr | This study |
| pUCyieO | 2.2-kb PstI-BamHI fragment containing yieP (hypothetical protein) and -O (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCyjiO | 2.8-kb SphI-BamHI fragment containing yjiO (putative transporter) and -N (hypothetical protein) genes cloned into pUC119, Apr | This study |
| Plasmids carrying MATE-type transporter ORFs | ||
| pUCydhE | 1.6-kb PstI-BamHI fragment containing ydhE (multidrug transporter) gene cloned into pUC119, Apr | This study |
| Plasmids carrying RND-type transporter ORFs | ||
| pUCacrAB | 6.0-kb PstI SmaI fragment containing acrR (regulator), -A (MFP), and -B (multidrug transporter) genes cloned into pUC119, Apr | This study |
| pUCcusA | 9.0-kb SphI-EcoRI fragment containing cusS (formerly ybcZ; sensor), cusR/C/F/B (ylcA/B/C/D: regulator/putative OMF/putative binding protein/putative MFP), and cusA (ybdE: putative metal ion or drug transporter) genes cloned into pUC119, Apr | This study |
| pUCyegMNOB | 14.2-kb SphI-EcoRI fragment containing yegK/L (putative two-component system), yegM/N/O/B (putative MFP/putative RND transporter/putative RND transporter/putative MF transporter), and baeS/R (putative two-component system) genes cloned into pUC119, Apr | This study |
| pUCacrD | 3.5-kb HindIII-SacI fragment containing acrD (drug transporter) gene cloned into pUC119, Apr | This study |
| pUCacrEF | 5.9-kb SphI-SalI fragment containing envR (regulator), acrE/F (MFP/multidrug transporter) genes cloned into pUC119, Apr | This study |
| pUCyhiUV | 4.7-kb PstI-BamHI fragment containing yhiU/V (putative MFP/putative transporter) genes cloned into pUC119, Apr | This study |
| Plasmids carrying SMR-type transporter ORFs | ||
| pUCemrE | 1.2-kb SmaI-BamHI fragment containing emrE (drug transporter) gene cloned into pUC118, Apr | This study |
| pUCydgFE | 1.1-kb SalI-BamHI fragment containing ydgF/E (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCsugE | 1.0-kb PstI-SmaI fragment containing sugE (putative transporter) gene cloned into pUC119, Apr | This study |
| Plasmids carrying ABC-type transporter ORFs | ||
| pUCmdlAB | 4.2-kb SphI-EcoRI fragment containing mdlA/B (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCybjYZ | 3.7-kb PstI-SalI fragment containing ybjY/Z (putative MFP/putative transporter) genes cloned into pUC119, Apr | This study |
| pUCyddA | 2.3-kb SalI-SacI fragment containing yddA (putative transporter) gene cloned into pUC119, Apr | This study |
| pUCyojIH | 4.0-kb SphI-SalI fragment containing yojI/H (putative transporter) genes cloned into pUC119, Apr | This study |
| pUCyhiH | 5.8-kb SphI-BamHI fragment containing yhiI/H (putative regulator/putative transporter) and yhhJ (hypothetical protein) genes cloned into pUC119, Apr | This study |
| Plasmids carrying drug transporter ORFs under control of trc promoter | ||
| MF type | ||
| pTrcHyajR | 1.4-kb NcoI-PstI fragment containing yajR gene cloned into pTrc6His, Apr | This study |
| pTrcHyceE | 1.2-kb NcoI-PstI fragment containing yceE gene cloned into pTrc6His, Apr | This study |
| pTrcHyceL | 1.2-kb NcoI-PstI fragment containing yceL gene cloned into pTrc6His, Apr | This study |
| pTrcHydeA | 1.2-kb NcoI-PstI fragment containing ydeA gene cloned into pTrc6His, Apr | This study |
| pTrcHydeF | 1.2-kb EcoRI-PstI fragment containing ydeF gene cloned into pTrc6His, Apr | This study |
| pTrcHynfM | 1.3-kb NcoI-PstI fragment containing ynfM gene cloned into pTrc6His, Apr | This study |
| pTrcHydhC | 1.2-kb NcoI-SalI fragment containing ydhC gene cloned into pTrc6His, Apr | This study |
| pTrcHydiM | 1.2-kb NcoI-SalI fragment containing ydiM gene cloned into pTrc6His, Apr | This study |
| pTrcHyebQ | 1.5-kb NcoI-SalI fragment containing yebQ gene cloned into pTrc6His, Apr | This study |
| pTrcHyegB | 1.4-kb NcoI-PstI fragment containing yegB gene cloned into pTrc6His, Apr | This study |
| pTrcHbcr | 1.2-kb NcoI-PstI fragment containing bcr gene cloned into pTrc6His, Apr | This study |
| pTrcHyidY | 1.2-kb NcoI-PstI fragment containing yidY gene cloned into pTrc6His, Apr | This study |
| pTrcHyieO | 1.4-kb NcoI-PstI fragment containing yieO gene cloned into pTrc6His, Apr | This study |
| pTrcHyjiO | 1.2-kb NcoI-PstI fragment containing yjiO gene cloned into pTrc6His, Apr | This study |
| RND type | ||
| pTrcHcusAB | 4.4-kb NcoI-XbaI fragment containing cusA and cusB genes cloned into pTrc6His, Apr | This study |
| pTrcHyegN | 3.1-kb NcoI-PstI fragment containing yegN gene cloned into pTrc6His, Apr | This study |
| pTrcHyegO | 3.1-kb EcoRI-PstI fragment containing yegO gene cloned into pTrc6His, Apr | This study |
| pTrcHacrD | 3.1-kb NcoI-SalI fragment containing acrD gene cloned into pTrc6His, Apr | This study |
| pTrcHacrEF | 4.3-kb SacI-PstI fragment containing acrE and acrF genes cloned into pTrc6His, Apr | This study |
| SMR type | ||
| pTrcHydgFE | 0.7-kb EcoRI-PstI fragment containing ydgF and ydgE genes cloned into pTrc6His, Apr | This study |
| pTrcHsugE | 0.5-kb NcoI-PstI fragment containing sugE gene cloned into pTrc6His, Apr | This study |
| ABC type | ||
| pTrcHmdlA | 1.8-kb EcoRI-PstI fragment containing mdlA gene cloned into pTrc6His, Apr | This study |
| pTrcHmdlB | 1.8-kb NcoI-PstI fragment containing mdlB gene cloned into pTrc6His, Apr | This study |
| pTrcHybjZ | 2.0-kb NcoI-PstI fragment containing ybjZ gene cloned into pTrc6His, Apr | This study |
| pTrcHyddA | 1.7-kb NcoI-PstI fragment containing yddA gene cloned into pTrc6His, Apr | This study |
| pTrcHyojI | 1.6-kb NcoI-PstI fragment containing yojI gene cloned into pTrc6His, Apr | This study |
| pTrcHyojH | 1.7-kb EcoRI-PstI fragment containing yojH gene cloned into pTrc6His, Apr | This study |
| pTrcHyhiH | 2.7-kb NcoI-PstI fragment containing yhiH gene cloned into pTrc6His, Apr | This study |
a
DNA fragments cloned in pUC vectors contained the endogenous promoter regions that were expected to work at the drug resistance measurements without induction.