TABLE 1.
Bacterial strains and plasmids used in this work
| Strain or plasmid | Characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. erythraea NRRL2338 | Used as parental strain, wild type | DSM 40517 |
| ΔpccD strain | S. erythraea pccD null mutant, thiostrepton resistance | This work |
| WT/PIB-pccD | pccD overexpression strain, WT carrying pIB-pccD | This work |
| E. coli Rosetta (DE3) | F− ompT hsdSB(rB− mB−) gal dcm λ(DE3)pRARE2(Camr) | Novagen |
| Plasmids | ||
| pET19b | Expression vector, Ampr | Novagen |
| pET-pccD | pET19b derivative carrying pccD | This work |
| pMD-18T | TA-cloning vector | TaKaRa |
| pUC19-tsr | pUC18 derivative containing a 1.36-kb fragment of a thiostrepton resistance cassette in the BamHI/SmaI sites | 34 |
| pUC-pccD | pUC19-tsr, with the 1.5-kb DNA fragments upstream and downstream of pccD gene inserted upstream and downstream of tsr correspondingly | This work |
| pIB139 | E. coli-S. erythraea integrative shuttle vector containing a strong constitutive ermE* promoter, apramycin resistance | 35 |
| pIB-pccD | pIB139 carrying an extra pccD for the gene overexpression | This work |