Table 3 Strains and plasmids used in this study.
Strains or plasmids | Description | Source/Reference |
|---|---|---|
S. tsukubaensis | ||
NRRL 18488 | Wild-type | |
ΔsigG1 | ΔsigG1::apr; AprR | This study |
ΔrsfG | ΔrsfG::apr; AprR | This study |
ΔsigG1-rsfG | Δ sigG1-rsfG::apr; AprR | This study |
WT pIJ12333 | Wild-type expressing pIJ12333 empty vector | This study |
ΔsigG1 pIJ12333 + sigG1 | ΔsigG1::apr expressing pIJ12333-sigG1; TsrR AprR (ɸC31-transduction) | This study |
ΔsigG1-rsfG1 pIJ12333 + sigG1 | ΔSTSU_11555-60::apr expressing pIJ12333-STSU_11560; TsrR AprR (ɸC31-transduction) | This study |
∆oxyR | ΔoxyR::apr; AprR | (Pires et al., unpublished results) |
E. coli | ||
BTH101 | F- cya-99 araD139 galE15 galK16 rpsL1 (Strr) hsdR2 mcrA1 mcrB1 | |
BW25113 [pIJ790] | Δ(araD-araB)567, ΔlacZ4787(::rrnB-4), lacIp-4000(lacIQ), λ-, rpoS369(Am), rph-1, Δ(rhaD-rhaB)568, hsdR514; CmR | |
DH5α | F- ɸ80lacZ∆M15 ∆(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-, mK +) phoA supE44 λ– thi-1 gyrA96 relA1 | |
ET12567 [pUZ8002] | dam, dcm, hsd, KanR , CmR | |
Nico21 | can::CBD fhuA2 [lon] ompT gal (λ DE3) [dcm] arnA::CBD slyD::CBD glmS6Ala ∆hsdS λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5 | New England Biolabs |
Plasmids | ||
pIJ12333 | Vector for conjugal transfer of DNA from E. coli to Streptomyces sp.; integrative (ɸC31-transduction), HygR TsrR | |
pIJ12333-sigG1 | pIJ12333 carrying sigG1 preceded by its native promoter, TsrR | This study |
pIJ773 | Plasmid template for amplification of the apr-oriT cassette for Redirect™ PCR-targeting | |
pIJ790 | Modified l RED recombination plasmid [oriR101] [repA101(ts)] araBp-gam-be-exo, CmR | |
pUZ8002 | RP4 derivative with defective oriT, KanR | |
pET15b | T7 expression vector, AmpR | Novagen |
pET15b-sigG1 | pET15b-sigG1 full length coding sequence | This study |
pRSFDuet-1™ | T7 co-expression vector. Contains two multiple cloning sites (MCS), KanR | Novagen |
pRSFDuet3 | pRSFDuet-1 carrying rsfG cloned into MCS1 and sigG1 cloned into MCS2 of pRSFDuet-1 | This study |
pUT18C | Two-hybrid vector, N-terminal cyAT18fusion, AmpR | |
pUT18 | Two-hybrid vector, C-terminal cyAT18fusion, AmpR | |
pKT25 | Two-hybrid vector, N-terminal cyAT25fusion, KanR | |
pKNT25 | Two-hybrid vector, C-terminal cyAT25fusion, KanR | |
pKT25-zip | Derivative of pKT25 carrying the leucine zipper of GCN4 fused in frame to T25 | |
pUT18C-zip | Derivative of pUT18C carrying the leucine zipper of GCN4 fused in frame to T18 | |
