TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Characteristic(s)a | Reference or source |
|---|---|---|
| Escherichia coli strains | ||
| DH5α | General cloning host | 68 |
| BW25113 | Strain for propagation of a recombination plasmid | 69 |
| ET12567/pUZ8002 | Strain for intergeneric conjugation | 70 |
| Streptomyces strains | ||
| S. iranensis HM 35 | Wild-type strain | 17 |
| S. iranensis ΔhutF | S. iranensis hutF::aac(3)IV mutant | This study |
| S. iranensis ΔrapK | S. iranensis rapK::aac(3)IV mutant | This study |
| Plasmids | ||
| pIJ790 | Helper plasmid, RED/ET recombination plasmid | 47 |
| pRSFDuet-1 | E. coli vector for cloning, Kanr | Novagen |
| pOJ260 | Suicide vector nonreplicating in Streptomyces, Aprr | 23 |
| pRSFDuet_pSG5 | Cloning vector, Kanr, pSG5 replicon | This study |
| pRSFDuet_pSG5_kan | E. coli-Streptomyces shuttle vector, cloning vector, Kanr, pSG5 replicon, oriT | This study |
| pKOSi | E. coli-Streptomyces shuttle vector, cloning vector, Kanr, pSG5 replicon | This study |
| pKOSi_hutF | Vector for disruption of hutF, based on pKOSi, Kanr | This study |
| pKOSi_rapK | Vector for disruption of rapK, based on pKOSi, Kanr | This study |
| pKOSi_ΔhutF | pKOSi_hutF::aac(3)IV Kanr Aprr | This study |
| pKOSi_ΔrapK | pKOSi_rapK::aac(3)IV Kanr Aprr | This study |
| pTNM | Source of pSG5 for pKOSi | 42 |
| pIJ773 | Template for amplification of aac(3)IV (Aprr) cassette + oriT | 47 |
a
Kanr, kanamycin resistance; Aprr, apramycin resistance.