Table 1.
Bacterial strains and plasmids used in this study
| Bacterial strain or plasmid | Relevant genotype or phenotype or description | Source or reference(s) |
|---|---|---|
| E. coli strains | ||
| DH5α | φ80dlacZΔM15 recA1 endA1 gyrAB thi-1 hsdR17(rK− mK+) supE44 relA1 deoR Δ(lacZYA-argF)U169 phoA | Promega |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) dcm gal λ(DE3) | Promega |
| BW25113/pIJ790 | Δ(araD-araB)567 ΔlacZ4787(::rrnB4) lacIp-40000(lacIq) λ−rpoS369(Am) rph-1 Δ(rhaD rhaB)568 hsdR514 on the bacterial chromosome; oriR101 repA1001(Ts) araBp-gam-be-exo on the pIJ790 plasmid | 11 |
| ET12567/pUZ8002 | dam dcm hsdS Cmr Tetr on the bacterial chromosome; tra KanrRP4 23 on pUZ8002 | 29 |
| US0 | ΔpyrF ΔhisB F′ | 25 |
| S. coelicolor strains | ||
| M145 | SCP1− SCP2− | 2 |
| M851 | M145 ΔadpASc | 36 |
| M145+pIJ6902-hyg | M145+pIJ6902-hyg | This study |
| M851+adpA(TTG) | M851+pIJ6902/2528-hyg | This study |
| M851+pIJ6902-hyg | M851+pIJ6902-hyg | This study |
| M851+LadpA(TTA) | M851+pHL71 | This study |
| M851+LadpA(TTG) | M851+pHL72 | This study |
| Plasmids | ||
| 2StC13 | Cosmid carrying adpASc gene | 33; http://strepdb.streptomyces.org.uk |
| pHZ2528 | pSET152 with 2.4-kb fragment harboring the adpASc gene (with TTG codon) | 36 |
| pHL71 | pSET152 with 3.2-kb fragment harboring the adpASc gene (with TTA codon) and surrounding genes SCO2791 and SCO2793 | 36 |
| pHL72 | pSET152 with 3.2-kb fragment harboring the adpASc gene (with TTG codon) and surounding genes SCO2791 and SCO2793 | 36 |
| pIJ6902/2528-hyg | pIJ6902-hyg containing adpASc under the control of the thiostrepton-inducible PtipA promoter | This study |
| pIJ6902-hyg | pIJ6902 derivative containing Hygr instead of Aprar | This study |
| pET-21a(+)adpAHis6 | pET-21a(+) harboring adpASc gene cloned into BamHI and XhoI sites | This study |
| pET-28a(+)His6-nuc-adpA(51) | pET28a(+) expressing His6-Nuc-AdpA(51) fusion protein cloned into BamHI and NotI restriction sites | This study |
| pB1H1-adpASc | pB1H1 vector harboring the adpASc gene cloned into restriction sites NotI and AvrII | This study |
| pH3U3-18random | pH3U3 vector harboring 18-bp randomized library cloned into NotI and SgsI restriction sites | This study |