TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant properties | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | (Δlac)U169 hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 40 |
| S17-1 | Smr, recA thi pro hsdRM+ RP4-2-Tc::Mu-Km::Tn7 λpir | 46 |
| BL21λ(DE3) pLysS | ompT hsdS(rB− mB−) dcm+endA galλ BL21(DE3) | Novagen |
| A. brasilense | ||
| Sp7 | Wild-type strain | 21 |
| Cd | Wild-type carotenoid-producing strain | 21 |
| Car-1 | Carotenoid-producing chrR:Tn5 mutant of A. brasilense Sp7 | 23 |
| Plasmids | ||
| pMMB206 | Broad-host-range, low-copy-number lactose-inducible expression vector; Cmr | 25 |
| pBBR1MCS-3 | Broad-host-range, medium-copy-number cloning vector; Tcr | 26 |
| pAK024 | egfp cloned into EcoRI-BamHI sites of pMMB206 | This work |
| pAK032 | pBBR1MCS-3 replicon-based lactose-inducible expression vector; Tcr | This work |
| pAK040 | pBBR1MCS-3 replicon-based lactose-inducible EGFP expressing plasmid; Tcr | This work |
| pAK041 | Codon-optimized A. annua ads cloned into NdeI-PstI sites of pAK032 | This work |
| pAK042 | E. coli ispA cloned into PstI-XhoI sites of pAK041 | This work |
| pAK043 | A. brasilense rpoE1 cloned into XhoI-KpnI sites of pAK042 | This work |
| pAK050 | As pAK042, but ispA replaced with its codon-optimized version | This work |
| pAK051 | As pAK043, but ispA replaced with its codon-optimized version | This work |
| pAK054 | crtE1 cloned into NdeI-HindIII sites of pET28a | This work |
| pAK055 | crtE2 cloned into NdeI-HindIII sites of pET28a | This work |
| pAK056 | Codon-optimized C. roseus gpps cloned into NdeI-XhoI sites of pAK032 | This work |
| pAK057 | Codon-optimized C. roseus ges cloned into NdeI-XhoI sites of pAK032 | This work |
| pAK058 | Codon-optimized C. roseus ges cloned into XhoI-KpnI sites of pAK056 | This work |
| pAK059 | As pAK058, but gpps replaced with mutated E. coli ispA | This work |