TABLE 1.
Streptomyces vectors and recombinant plasmids
| Plasmids | Relevant characteristics and constructiona | Reference |
|---|---|---|
| pIJ486 | High-copy-number cloning vector, tsr | 47 |
| pGM9 | pSG5-derived temperature-sensitive replication vector, tsr aphII | 35 |
| pIJ941 | Low-copy-number SCP2* derivative plasmid, tsr hyg | 25 |
| pIJ2314 | BamHI fragment (positions 13–14) cloned in pIJ922, harboring the wild-type actIII gene, tsr | 32 |
| pAM73 | pGM9 derivative obtained from S. lividans TK21::pSCNB06A chromosomal DNA partially digested with Sau3AI, followed by ligation and transformation of S. lividans | This work |
| pAM75 | pGM9 derivative obtained from S. coelicolor J1501::pSCNB06A chromosomal DNA partially digested with Sau3AI, followed by ligation and transformation of S. lividans | This work |
| pSCNB01b | SacII (2274)-XhoI (3465) fragment cloned in pGM9 (EcoRI-HindIII sites); contains the 3′ truncated orf10 gene | This work |
| pSCNB03b | PstI (1881)-XhoI (3465) fragment cloned in pGM9 (EcoRI-HindIII sites); contains orf10 gene | This work |
| pSCNB04b | PstI (1881)-BamHI (4847) fragment cloned in pGM9 (EcoRI-HindIII sites); contains orf10 and orf11 genes | This work |
| pSCNB05c | NcoI (3094)-BamHI (4847) fragment cloned in pGM9 (BglII site); contains orf11 gene | This work |
| pSCNB06Ac | SacII (2274–2674) fragment containing an internal region of orf10 gene cloned in pGM9 (EcoRI-HindIII sites) | This work |
| pSCNB06Bd | pSCNB06A with the internal region of orf10 gene cloned in the opposite orientation | This work |
| pSCNB07c | SmaI (809)-BglII (2389) and NruI (3049)-BamHI (4847) fragments cloned in the same direction in pGM9 (EcoRI-HindIII sites) | This work |
| pSCNB08Bc | SmaI (809)-BamHI (2152) and NcoI (3094)-BamHI (4847) fragments cloned in the same direction in pGM9 (EcoRI-HindIII sites) | This work |
| pSCNB09c | NruI (3049)-BamHI (4847) fragment cloned in pGM9 (EcoRI-HindIII sites); contains orf11 gene | This work |
| pSCNB010d | SacII (2274)-BamHI (4847) fragment cloned in pGM9 (EcoRI-HindIII sites); contains the 3′-truncated orf10 and entire orf11 genes | This work |
| pSCNB013b | PstI (1881)-XhoI (3465) fragment cloned in pIJ941 (EcoRV site) | This work |
| pSCNB014b | PstI (1881)-XhoI (3465) fragment cloned in pIJ486 (EcoRI-HindIII sites) | This work |
tsr, thiostrepton resistance gene; aphII, aminoglycoside phosphotransferase gene from Tn5; hyg, hygromycin resistance gene. Numbers in parentheses after a restriction enzyme indicate nucleotide positions.
Constructed either directly or as blunt-ended fragments through end filling with the Klenow fragment of DNA polymerase I or by T4 DNA polymerase treatment in the intermediate E. coli vector pUC19, rescued, and ligated to Streptomyces vectors as indicated.
Constructed either directly or as blunt-ended fragments through end filling with the Klenow fragment of DNA polymerase I or by T4 DNA polymerase treatment in the intermediate E. coli vector pU2925, then rescued, and ligated to Streptomyces vectors as indicated.
Constructed either directly or as blunt-ended fragments through end filling with the Klenow fragment of DNA polymerase I or by T4 DNA polymerase treatment in an intermediate E. coli vector pSU21, then rescued, and ligated to Streptomyces vectors as indicated.