TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotype/comments | Source/reference |
|---|---|---|
| Strains | ||
| S. coelicolor A3(2) | ||
| M600 | SCP1− SCP2− | 21 |
| J2130 | ΔsigE SCP1− SCP2− | 27 |
| J2142 | ΔcseB::hyg SCP1− SCP2− | 28 |
| J2172 | ΔcseA SCP1− SCP2− | This study |
| E. coli | ||
| ET12567(pUZ8002) | ET12567 containing helper plasmid pUZ8002 | 27 |
| Plasmids | ||
| pGEX-KG | E. coli overexpression vector | 11 |
| pKC1132 | Conjugative vector for gene disruption in Streptomyces (Aprr) | 5 |
| pSET152 | φC31 attP-int-derived integration vector for the conjugal transfer of DNA from E. coli to Streptomyces spp. (Aprr) | 5 |
| pIJ486 | Multicopy Streptomyces promoter-probe plasmid containing neo as reporter gene (Thior) | 39 |
| pIJ6880 | 0.75-kb sigE promoter region (BglII) in pIJ486 (BamHI) | 14 |
| pIJ6809 | pKC1132 carrying an in-frame cseA deletion allele | This study |
| pIJ6810 | pSET152 carrying the cseA gene driven by its native promoter (sigEp) | This study |
| pIJ6946 | pGEX-KG derivative overexpressing CseA lacking the first 44 amino acids | This study |
| pIJ6962 | pSET152 carrying the cseA gene driven by its native promoter (sigEp) | This study |
| pIJ6963 | As pIJ6962, but carrying the TTG1-to-CTC mutation | This study |
| pIJ6964 | As pIJ6962, but with a TGA stop codon introduced between TTG1 and GTG2 | This study |
| pIJ6065 | As pIJ6962, but carrying the GTG2-to-GTC mutation | This study |
| pIJ6966 | As pIJ6962, but carrying the GTG3-to-GTC mutation | This study |
| pIJ6967 | As pIJ6962, but carrying the C37A substitution | This study |