Table 2 Strains, plasmids, and primers used in this study
Strains, plasmids, and primers | Description | Reference |
|---|---|---|
Bacterial strains | Â | Â |
 Streptomyces sp. 139 | Ebosin producing strain | Our lab |
 Streptomyces sp. 139 D2 | ste2 knockout mutant of Streptomyces sp.139 | This study |
 Streptomyces sp. 139 C2 | ste2-complemented strain | This study |
 E. coli DH5α | F− recA1 endA1 hsdR17 deoR thi-1 supE44 gyrA96 relA1 Δ(lacZYA- argF) U169λ−(φ80dlacZΔM15) |  |
 E. coli ET12567 | methylation-deficient E. coli; dam− dcm− hsdM | 44 |
 E. coli BL21 (DE3) | F− ompT hsdSB(rB− mB−) dcm gal λ(DE3) | Novagen |
Plasmids | Â | Â |
 pKC1139 | Shuttle plasmid (E.coli–Streptomyces); pSG5, pBR322; aac[3]IV lacZa oriTRK2; Amr | 43 |
 pKC2D | pKC1139 derived plasmid carrying F1, F2 and Kmr fragments; Kmr Amr | This study |
 pKC2C | pKC1139 derived plasmid carrying 0.45-kb ErmE* promoter fragment and ste2; Amr | This study |
 pGEM-3Zf-ErmE* | Resource of ErmE* promoter; Apr | 45 |
 pET30a | T7 promoter, His-tag; Kmr | Novagen |
 pET30a-ste2 | pET30a derived plasmid carrying ste2; Kmr | This study |
Primers | Â | Â |
 P1 (EcoRI) | CTGGAATTCGTGCCCTTGCCCTGGAT | F1 |
 P2 (XbaI) | GCCTCTAGACTTCGCCTTGGTCTTCT | F1 |
 P3 (XbaI) | GCATCTAGAGGCAGCCAGAAGCAGGAAC | F2 |
 P4 (HindIII) | AGCAAGCTTACAGGATGGAGCGGAGG | F2 |
 P5 (BamHI) | CGCGGATCCATGGGTGCGCAAGGCATT | ste2 |
 P6 (HindIII) | CCCAAGCTTTTACTGCTGCTGCGCCAG | ste2 |
 ste5 forward | GCTGATCCTGCTGGTGGTGC |  |
 ste5 reverse | CCATCGTGCGGAACTTGAGG |  |
 ste8 forward | CTCGGCAAGCTCAGCCAGAC |  |
 ste8 reverse | CGAGCAGCAGGAACAGCACC |  |
 ste17 forward | CTGGACGGCGACGAGAT |  |
 ste17 reverse | CGACGCAGTGGAACGAG |  |
 hrdB forward | TGGTCGAGGTCATCAACAAG |  |
 hrdB reverse | TGGACCTCGATGACCTTCTC |  |