TABLE 2.
Plasmids used in this study
| Plasmid | Relevant characteristics and/or use | Resistance | Source and/or reference(s) |
|---|---|---|---|
| pUZ8002 | Encodes transfer functions for mobilization of oriT-containing vectors from E. coli to Streptomyces by conjugative transfer | Kanamycin | M. Buttner, John Innes Centre (31) |
| pIJ6902 | Expression vector that contains thiostrepton-inducible ptipA promoter; integrates into the φC31 attB site in S. coelicolor | Apramycin | John Innes Centre (24) |
| pAU316 | pIJ6902 harboring bldG; for expression of BldG in S. coelicolor | Apramycin | This study |
| pAU317 | pIJ6902 harboring sco3548; for expression of SCO3548 in S. coelicolor | Apramycin | This study |
| pAU318 | pET30(a)+ derivative containing sco3548 cloned downstream of a 10-histidine tag; for overexpression of His10-SCO3548 in E. coli | Kanamycin | This study |
| pAU365 | pIJ6902 derivative containing the entire bldG and sco3548 coding regions and their native intergenic region; for expression of BldG and SCO3548 in S. coelicolor | Apramycin | This study |
| pGEX-2TK | Expression vector that contains IPTG-inducible tac promoter; for overexpression of protein with an N-terminal GST tag in E. coli | Ampicillin | Amersham Biosciences |
| pAU375 | pGEX-2TK harboring bldG; for overexpression of GST-BldG in E. coli | Ampicillin | This study |
| pAU376 | pGEX-2TK harboring sco3548; for overexpression of GST-SCO3548 in E. coli | Ampicillin | This study |
| pKT25 | pSU40 derivative that carries the T25 fragment of B. pertussis adenylate cyclase (residues 1 to 224) with a multicloning sequence at the 3′ end of T25 | Kanamycin | 28, 30 |
| pUT18 | pUC19 derivative that carries the T18 fragment of B. pertussis adenylate cyclase (residues 225 to 399) with a multicloning sequence at the 5′ end of T18 | Ampicillin | 28 |
| pUT18C | pUC19 derivative that carries the T18 fragment of B. pertussis adenylate cyclase with a multicloning sequence at the 3′ end of T18 | Ampicillin | 28 |
| pKT25Zip | pKT25 harboring the T25 fragment fused in frame to the 35-codon leucine zipper domain of the yeast GNC4 activator | Kanamycin | 30 |
| pUT18CZip | pUT18C harboring the 35-codon leucine zipper domain of the yeast GNC4 activator fused in frame to the T18 fragment | Ampicillin | 28 |
| pAU377 | pKT25 harboring bldG; for expression of T25-BldG in BTH101 | Kanamycin | This study |
| pAU378 | pUT18 harboring bldG; for expression of BldG-T18 in BTH101 | Ampicillin | This study |
| pAU379 | pUT18C harboring bldG; for expression of T18-BldG in BTH101 | Ampicillin | This study |
| pAU380 | pKT25 harboring sco3548; for expression of T25-SCO3548 in BTH101 | Kanamycin | This study |
| pAU381 | pUT18 harboring sco3548; for expression of SCO3548-T18 in BTH101 | Ampicillin | This study |
| pAU382 | pUT18C harboring sco3548; for expression of T18-SCO3548 in BTH101 | Ampicillin | This study |