TABLE 1.
Bacterial strains, plasmids, and libraries
| Bacterial strain, plasmid, or construct | Relevant genotype or description | Reference |
|---|---|---|
| Strains | ||
| E. coli JM109 | F′ traD36 proA+ proB+ lacIq Δ(lacZ)M15 Δ(lac-proAB) glnV44 gyrA96 recA1 relA1 endA1 thi hsdR17 | 22 |
| E. coli ET12567 | dam dcm hsdM hsdS hsdR cat tet | 18 |
| S. coelicolor M145 | Wild type (SCP1− SCP2−) | 15 |
| S. coelicolor GSA3 | M145 ΔssgA(::aadA) | 28 |
| Plasmids and constructs | ||
| pIJ2925 | Derivative of pUC19 with BglII sites flanking the multiple cloning site | 10 |
| pHJL401 | Shuttle vector with ori for maintenance in E. coli (pUC) and ori for replication in Streptomyces (SCP2*) | 16 |
| pGWS32 | pHJL401 with a 2-kb fragment harboring the ssgR and ssgA gene cluster (including promoter regions) | This study |
| pGWS278 (library) | pIJ2925 with the random mutagenic PCR product of ssgA (reaction with additional dGTP) | This study |
| pGWS279 (library) | pIJ2925 with the random mutagenic PCR product of ssgA (reaction with additional dTTP) | This study |
| pGWS280 (library) | pGWS32 with the wild-type ssgA sequence from the BamHI site replaced with inserts from pGWS278 and pGWS279 | This study |