TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| S. coelicolor | ||
| M145 | Parental strain SCP1− SCP2− | 22 |
| T124 | M145 (accB/pTR124); Thr Hygr | This work |
| T149 | T124 containing pTR149 integrated in the att site of φC31; Thr Hygr Amr | This work |
| T149A | T149 with the wild-type accB copy of the chromosome replaced by the accB::hyg mutant allele; Hygr Amr | This work |
| E. coli | ||
| DH5α | ΔlacU169 (φ80lacZΔM15) endA1 recA1 hsdR17 deoR supE44 thi-1 λ− gyrA96 relA1 | 15 |
| BL21λ(DE3) | ompT(DE3) | 43 |
| ET 12567 | supE44 hsdS20 ara-14 proA2 lacY galK2 rpsL20 xyl-5 mtl-1 Δdam Δdcm ΔhsdM Cmr | 29 |
| RG7 | DH5α carrying pCL1 and pBA11 plasmids | 43 |
| Plasmids | ||
| pBluescript SK(+) | Phagemid vector (AprlacZ′) | Stratagene |
| PCR-Blunt | Used for cloning PCR products | Invitrogen |
| pGEM-T Easy | Used for cloning PCR products | Promega |
| pIJ2925 | pUC18 derivative (AprlacZ′) | 22 |
| pSET151 | Used for the conjugal transfer of DNA from E. coli to Streptomyces spp. (Apr ThrlacZ′) | 3 |
| pET22b(+) | Phagemid vector (AprlacZ′) for expression of recombinant proteins under control of strong T7 transcription and translation signals | Novagen |
| pUZ8002 | RK2 derivative with defective oriT (Kmr) | 31 |
| pIJ8600 | Used for the conjugal transfer of DNA from E. coli to Streptomyces spp. and for expression of recombinant proteins under tipA promoter | 44 |
| pBA11 | Vector containing E. coli birA gene | 1 |
| pTR45 | pSK(+) with a chromosomal Pstl insert carrying accA2 | 36 |
| pRM08 | pSK(+) with an Sstl insert carrying accBE | This work |
| pTR82 | pSK(+) carrying accBE with an NdeI site in the translation start site of accB | This work |
| pTR88 | pET22b(+) with accBE under control of strong T7 transcription and translation signals | This work |
| pTR90 | pET22b(+) with accB under control of strong T7 transcription and translation signals | This work |
| pTR94 | pIJ8600 derivative with a deletion of the int and att sites and carrying the accBE genes under tipAp control | This work |
| pTR107 | pET22b(+) with accE under control of strong T7 transcription and translation signals | This work |
| pTR124 | pSET151 with a hyg (Hygr) gene inserted in the accB coding region | This work |
| pTR141 | pIJ8600 derivative carrying oriT RK2, ori pUC18, attP site, int φC31, and aac(3)IV (Amr) | This work |
| pTR149 | pTR141 with a KpnI insert carrying accBE | This work |
| pTR204 | pET21a(+) with accA2 under control of strong T7 transcription and translation signals | This work |
| pTR237 | pET28a(+) with an accE His tag fusion gene | This work |