TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Bacterial strains | ||
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17(rK− mK+) deoR thi-1,supE44 gyrA96 relA1 λ−phoA | Toyobo |
| BL21(DE3) | F−ompT hsdS(rB− mB−) gal dcm λ (DE3) | Novagen |
| P. putida | ||
| KT2440 | Naturally Cmr | 2 |
| KT2440(pCAR1) | KT2440 harboring pCAR1 | 31 |
| KT2440(pCAR1Δpmr) | KT2440 harboring pCAR1 carrying disrupted pmr gene by Gmr gene cassette | This study |
| KT2440(pCAR1pmrHis) | KT2440(pCAR1) carrying gene encoding His-tagged Pmr | This study |
| Plasmids | ||
| pET-26b(+) | Kmr, T7 promoter, lacI, pBR322 replicon | Novagen |
| pET-C-His-pmr | pET-26b(+), NdeI-XhoI fragment containing pmr | This study |
| pFLAG-CTC | Apr, lacI, tac promoter expressing FLAG tag | Sigma |
| pFLAG0017 | pFLAG-CTC, NdeI-SalI fragment containing PP_0017 | This study |
| pFLAG1366 | pFLAG-CTC, NdeI-SalI fragment containing PP_1366 | This study |
| pFLAG2947 | pFLAG-CTC, NdeI-SalI fragment containing PP_2947 | This study |
| pFLAG3693 | pFLAG-CTC, NdeI-SalI fragment containing PP_3693 | This study |
| pFLAG3765 | pFLAG-CTC, NdeI-SalI fragment containing PP_3765 | This study |
| pFLAGpmr | pFLAG-CTC, NdeI-SalI fragment containing pmr | This study |
| pFLP2 | Apr, ori1600 oriT(RP4), Flp recombinase expression vector | 16 |
| pFLP2Km | pFLP2, Kmr gene cassette inserted into its ScaI site | This study |
| pK19mobsacB | Kmr, oriT(RP4) sacB lacZα, pMB1 replicon | 29 |
| pK19mobsacBpmrHis | pK19mobsacB containing gene encoding Pmr with 6× His at its C-terminal end with Gmr gene cassette and FRT site at its 3′-terminal end | This study |
| pK19mobsacBpmrGm | pK19mobsacB, containing 3.8-kb EcoRI-PstI fragment including Gmr gene cassette inserted pmr flanking region (77486-77909 region of pCAR1 was replaced by Gmr gene) | This study |
| pSJ12 | pBluescript II KS(−) with 0.7-kb SmaI fragment containing a nonpolar Gmr gene cassette | 17 |
| pT7Blue T-vector | Apr, lacZα, T7 promoter, f1 origin, pUC/M13 priming sites | Novagen |
| pTKm | pT7Blue T-vector, Kmr gene cassette | 43 |
| pUC18 | Apr, lacZα, pMB9 replicon | 28 |
| pUB11 | pUC18 containing 75211-80781 region of pCAR1 | This study |