TABLE 1.
Plasmids and strains used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| S. avermitilis | ||
| ATCC 31267 | Wild-type strain (WT) | Laboratory stock |
| accD1::pKC1139 | accD1 disruption strain by insertion of pKC1139 | This study |
| DaccR | accR deletion mutant of strain ATCC 31267 | This study |
| CaccR | DaccR with insertion of plasmid pSET-CaccR | This study |
| A8 | Industrial high-yield avermectin-producing strain | Qilu Pharmaceutical |
| A8/DaccR | accR deletion mutant of strain A8 | This study |
| WT(pKC-ermE-accD1A1) | accD1A1 overexpression strain based on WT | This study |
| DaccR(pKC-ermE-accD1A1) | accD1A1 overexpression strain based on DaccR | This study |
| E. coli | ||
| JM109 | Host for DNA cloning | Laboratory stock |
| BL21(DE3) | Host for protein overexpression | Laboratory stock |
| Plasmids | ||
| pKC1139 | Temperature-sensitive shuttle vector for E. coli-Streptomyces | 41 |
| pSET152 | Integrated shuttle vector in Streptomyces | 41 |
| pET-28a(+) | Expression vector for His6-tagged protein purification in E. coli | Novagen |
| pKC-DaccD1 | accD1 disruption plasmid based on pKC1139 | This study |
| pKC-DaccR | accR deletion plasmid based on pKC1139 | This study |
| pSET-CaccR | accR complementation vector based on pSET152 | This study |
| pET-AccR | accR overexpression vector based on pET-28a(+) | This study |
| pKC-ermE-accD1A1 | accD1A1 overexpression vector based on pKC1139 carrying promoter ermE*p | This study |