TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | ΔlacU169 hsdR17 recA1 endA1 gyrA96 thiL relA1 | Gibco-BRL |
| S17-1 | Smr recA thi pro hsdR RP4-2(Tc::Mu Km::Tn7) | 24 |
| A. brasilense | ||
| Sp7 | Wild-type strain | 15 |
| Cd | Wild-type carotenoid-producing strain | 15 |
| Car-1 | Carotenoid-producing chrR1::Tn5 mutant of A. brasilense Sp7 | 16 |
| rpoE1::Km mutant | A. brasilense Sp7 rpoE1 gene disrupted by insertion of Kmr gene cassette | 25 |
| rpoH2::Km mutant | A. brasilense Sp7 rpoH2 gene disrupted by insertion of Kmr gene cassette | 19 |
| crtE1::Km mutant | A. brasilense Cd crtE1 gene disrupted by insertion of Kmr gene cassette | This work |
| crtE2::Km mutant | A. brasilense Cd crtE2 gene disrupted by insertion of Kmr gene cassette | This work |
| Plasmids | ||
| pSUP202 | ColE1 replicon, mobilizable, suicide vector for A. brasilense; Apr Cmr Tcr | 24 |
| pUC4K | Vector containing kanamycin resistance gene cassette | GE Healthcare |
| pAPD1 | crtE1::Km in pSUP202 | This work |
| pAPD2 | crtE2::Km in pSUP202 | This work |
| pCZ750 | Tetr Ampr; lacZ fusion reporter vector | 27 |
| pAPD3 | crtE1 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAPD4 | crtE2 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR1 | crtE2 promoter (deleted −35) region of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR2 | rpoH1 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR3 | rpoH2 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR4 | rpoH2 promoter −35 region (TGA to ACT) of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR5 | rpoH2 promoter −10 region (CG to GC) of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR6 | rpoH2 promoter (deleted −35) region of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR7 | rpoH3 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR8 | rpoH4 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pAKR9 | rpoH5 promoter sequence of A. brasilense Sp7 fused with lacZ in pCZ750 | This work |
| pMMB206 | Cmr; broad-host-range, low-copy-number expression vector | 26 |
| pAT5 | rpoE1 gene of A. brasilense Sp7 cloned in pMMB206 vector | 16 |
| pNG6 | rpoE2 gene of A. brasilense Sp7 cloned in pMMB206 vector | 17 |
| pAKR9 | rpoH1 gene of A. brasilense Sp7 cloned at EcoRI and PstI in pMMB206 vector | This work |
| pSNK10 | rpoH2 gene of A. brasilense Sp7 cloned in pMMB206 vector | 19 |
| pAKR11 | rpoH3 gene of A. brasilense Sp7 cloned in pMMB206 vector | This work |
| pAKR12 | rpoH4 gene of A. brasilense Sp7 cloned in pMMB206 vector | This work |
| pAKR13 | rpoH5 gene of A. brasilense Sp7 cloned in pMMB206 vector | This work |