TABLE 2.
Bacterial strains and plasmids used in this studya
| Strain or plasmid | Relevant phenotype or genotypeb | Reference or origin |
|---|---|---|
| E. coli strains | ||
| TG1 | supE hsd Δ5 thi Δ[lac-proAB] F′ [traD36 proAB lacIqlacZ ΔM15] | 28 |
| KAM3 | Derivative of TG1 that lacks acrB | 14 |
| TG1ΔtolC | Derivative of TG1 that lacks tolC | 16 |
| DH5α | recA endA1 hsdR17 supE4 gyrA96 relA1 Δ(lacZYA-argF)U169 (φ80dlacZΔ M15) | 28 |
| Plasmids | ||
| General | ||
| pHSG398 | Vector; CPr; multiple cloning site in lacZ | Takara Bio Inc. |
| pHSG399 | Vector; CPr; multiple cloning site in lacZ | Takara Bio Inc. |
| Plasmids carrying RND-type transporter ORFs | ||
| pHSGacrAB | 5.1-kb BamHI-SphI fragment containing acrA/B (MFP/multidrug transporter) genes cloned into pHSG398, CPr | This study |
| pHSGacrD | 3.5-kb HindIII-SacI fragment containing acrD (multidrug transporter) gene cloned into pHSG399, CPr | This study |
| pHSGacrEF | 5.0-kb SphI-SalI fragment containing acrE/F (MFP/multidrug transporter) genes cloned into pHSG399, CPr | This study |
| pHSGmdtABC | 7.7-kb SphI-EcoRI fragment containing mdtA/B/C (MFP/multidrug transporter/multidrug transporter) genes cloned into pHSG399, CPr | This study |
| pHSGmdtEF | 4.7-kb PstI-BamHI fragment containing mdtE/F (formerly named yhiU/V) (MFP/multidrug transporter) genes cloned into pHSG399, CPr | This study |
| Plasmids carrying MFS-type transporter ORFs | ||
| pHSGemrAB | 3.9-kb SalI-BamHI fragment containing emrR/A/B (regulator/MFP/multidrug transporter) genes cloned into pHSG399, CPr | This study |
| pHSGemrKY | 2.8-kb SphI-BamHI fragment containing emrK/Y (MFP/bile acid transporter) genes cloned into pHSG399, CPr | This study |
| pHSGmdfA | 1.6-kb KpnI-HindIII fragment containing mdfA (multidrug transporter) gene cloned into pHSG398, CPr | This study |
| Plasmid carrying ABC-type transporter ORFs | ||
| pHSGmacAB | 3.7-kb Pstl-SalI fragment containing macA/B (MFP/macrolide-specific transporter) genes cloned into pHSG399, CPr | This study |
| Plasmid carrying SMR-type transporter ORFs | ||
| pHSGemrE | 1.2-kb SmaI-BamHI fragment containing emrE (multidrug transporter) gene cloned into pHSG398, CPr | This study |
| Plasmid carrying MATE-type transporter ORFs | ||
| pHSGmdtK | 1.6-kb PstI-BamHI fragment containing mdtK (formerly named ydhE) (multidrug transporter) gene cloned into pHSG399, CPr | This study |
a
Gene fragments were cloned into pHSG vectors in the same orientation as the lactose promoter.
b
CP, chloramphenicol; MFP, membrane fusion protein.