TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Relevant characteristics or use | Source or reference |
|---|---|---|
| Strains | ||
| S. erythraea NRRL23338 | Parental strain, wild type | DSM 40517 |
| E. coli DH5α | F− ϕ80dlacZΔM (lacZYA-argF)U169 deoR | Invitrogen |
| E. coli BL21(DE3)-7101 | The strain for expression of GlnR | Lab stock |
| E. coli BL21(DE3) | F′ ompT rB− mB− (λDE3) | Invitrogen |
| S. erythraea phoPO | Overexpression of phoP; NRRL23338 integrated with pIB139-6965 | This study |
| S. erythraea ΔglnR | NRRL23338 glnR::tsr (glnR-null mutant) | Lab stock |
| S. erythraea null mutant | Plasmid-alone negative control strain with pIB139 integrated into the genome | This study |
| S. erythraea ΔglnR/pIB-glnR (glnRC) | glnR-complemented strain; ΔglnR strain carrying pIB-glnR | Lab stock |
| Plasmids | ||
| pET28a(+) | Vector with T7 RNA polymerase-based promoter for expression in E. coli BL21; hexahistidine tag with thrombin cleavage | Thermo Scientific |
| p7101 | pET28a(+) with glnR (SACE-7101) inserted in the NcoI-HindIII site | This study |
| p6965 | pET28a(+) with phoP (SACE-6965) inserted in the NcoI-HindIII site | This study |
| p6965130 | pET28a(+) with phoPDBD (domain for DNA binding, 390 bp of C terminus) inserted in NcoI-HindIII | This study |
| pUC18-tsr | pUC18 with tsr gene for thiostrepton resistance inserted into the BamHI-SmaI site | 32 |
| pIB139 | pSET152 with integrase of phiC31 and PermE, the strong promoter of Streptomyces | 32 |
| pIB139-7101 | pIB139 with glnR inserted into NdeI-EcoRV | This study |
| pIB139-6965 | pIB139 with phoP inserted into NdeI-EcoRV | This study |