TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| S. avermitilis | ||
| ATCC 31267 | Wild-type (WT) strain | Laboratory stock |
| A229 | Industrial strain | Qilu Pharmaceutical |
| ΔhspR | hspR deletion mutant | This study |
| ChspR | hspR complemented strain | This study |
| OhspR | hspR overexpression strain | This study |
| ΔphoP | phoP deletion mutant | 38 |
| ΔhspR/hspR-3FLAG | hspR complemented strain with HspR-3FLAG | This study |
| ΔphoP/hspR-3FLAG | phoP deletion mutant with HspR-3FLAG | This study |
| WT/hspR-3FLAG | WT strain with HspR-3FLAG | This study |
| WT/pKC1139 | WT strain carrying empty vector pKC1139 | This study |
| WT/pSET152 | WT strain carrying empty vector pSET152 | This study |
| OhspR/A229 | hspR overexpression strain based on A229 | This study |
| E. coli | ||
| JM109 | General cloning host for plasmid manipulation | Laboratory stock |
| BL21(DE3) | Host for protein expression | Novagen |
| Plasmids | ||
| pKC1139 | Multiple-copy temperature-sensitive E. coli-Streptomyces shuttle vector | 46 |
| pSET152 | Integrative E. coli-Streptomyces shuttle vector | 46 |
| pET-28a(+) | Vector for His6-tagged protein expression in E. coli | Novagen |
| pGEX-4T-1 | Vector for GST-tagged protein expression in E. coli | GE Healthcare |
| pCIMt005 | Multiple-copy temperature-sensitive E. coli-Streptomyces shuttle vector | 47 |
| pJL117 | pIJ2925 derivative carrying the Streptomyces strong constitutive promoter ermE*p | 49 |
| pKCΔhspR | hspR deletion vector based on pKC1139 | This study |
| pΔhspR | hspR deletion vector based on pCIMt005 | This study |
| pSET152-hspR | hspR complemented vector based on pSET152 | This study |
| pKC-erm-hspR | hspR overexpression vector based on pKC1139 | This study |
| pSET152-hspR- | hspR complemented vector with 3×FLAG-tagged | This study |
| 3FLAG | HspR on pSET152 | |
| pIJ10500 | Vector carrying 3×FLAG fragment | 50 |
| pET28-hspR | His6-HspR expression vector based on pET-28a(+) | This study |
| pGEX-phoP | GST-PhoP expression vector based on pGEX-4T-1 | 38 |