TABLE 1.
Bacterial strains and plasmids used or constructed in this study
| Strain or plasmid | Descriptiona | Source or referenceb |
|---|---|---|
| Strains | ||
| E. coli | ||
| JM109 | Strain used as a host for cloning and bioassay tester organism | ATCC |
| ET12567 | Strain harboring pUZ8002 plasmid; used for conjugating plasmid into Streptomyces | 24 |
| S. griseus | ||
| ATCC 700974 | Wild type | ATCC |
| SCAK1 | WT-derived strain containing pSET and hrdBp-metYXSO at attB | This study |
| SCAK2 | WT-derived strain containing pSET and hrdBp-abmD at attB | This study |
| SCAK3 | WT-derived strain containing pSET and hrdBp-metYXSO-hrdBp-abmD-hrdBp-malY-metB at attB | This study |
| SCAK4 | WT-derived strain containing pSET and hrdBp-ask-asd at attB | This study |
| SCAK5 | WT-derived strain containing pSETand hrdBp-metYXSO-hrdBp-ask-asd at attB | This study |
| S. albulus | ||
| NBRC 14147 | Genome used to amplify the genes for feedback-insensitive Ask and Asd | NBRC |
| Plasmids | ||
| pDrive | Used for subcloning (Kanr Ampr) | Qiagen |
| pSET(GUS) | Used for subcloning (Aprr) | 25 |
| pSE34-oriT | Multicopy plasmid in Streptomyces (Tsr) | 19 |
| pA10 | pSE34 derivative control | 19 |
| pCK1 | pSE34 derivative for ermE*p expression of metYXSO | This study |
| pCK2 | pSE34 derivative for ermE*p expression of malY-metB | This study |
| pCK3 | pSE34 derivative for ermE*p expression of mtcC-mtcB | This study |
| pCK4 | pSE34 derivative for ermE*p expression of sahH | This study |
| pCK5 | pSE34 derivative for ermE*p expression of metK-sahH | This study |
| pCK6 | pSE34 derivative for ermE*p expression of metE | This study |
| pCK7 | pSE34 derivative for ermE*p expression of metK-sahH-metE | This study |
a
Tsr, thiostrepton resistant; ermE*p, mutated ermE promoter.
b
ATCC, American Type Culture Collection; NBRC, Japanese NITE Biological Resource Center.