Table 1.
Bacterial strains, plasmids, cosmids, and primers used in this study.
| Strain, plasmid, cosmid | Description | Reference |
|---|---|---|
| S.coelicolor M145 | SCP1− SCP2−, reference strain | Kieser, 2000 |
| S.coelicolor ΔSCO4117 | SCO4117 replaced with acc(3)IV, AprR | This study |
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL endA1 nupG | Invitrogen |
| E. coli ET12567 | dam-13::Tn9, dcm-6, hsdM, hsdR | MacNeil et al., 1992 |
| E. coli ET12567/pUZ8002 | E. coli ET12567 harboring pUZ8002, a not self-transmissible plasmid which can mobilize oriT-containing plasmids by conjugation | Flett et al., 1997 |
| Bacillus subitilis | Indicator microorganism for CDA bioassay | |
| PLASMIDS AND COSMIDS | ||
| pMS82 | Integrative and conjugative vector, HygR | Gregory et al., 2003 |
| PCR™-Blunt II-TOPO® | Zero Blunt® TOPO® PCR Cloning Kit, KanR | Invitrogen |
| pIJ773 | Apr cassette in pIJ699 | Gust et al., 2003 |
| pΔSCO4117 | SCO4117 deletion construction, a Streptomyces non-replicative plasmid transmissible by conjugation, AprR KanR | This study |
| pMS82-SCO4117 | Integrative and conjugative plasmid derived from pMS82 with the completed SCO4117 gen under its own promoter control. HygR | This study |
| pCRISPR-Cas9 | Conjugative and thermosensitive plasmid harboring Cas9 | Tong et al., 2015 |
| pCRISPR-120 | pCRISPR-Cas9 harboring the apramycin resistance target sequence | This study |
| pCRISPR-SCO4117 | pCRISPR-120 harboring a 3.7 kb DNA fragment including SCO4117 | This study |
| pCRISPR-SgSCO4117 | pCRISPR-Cas9 harboring the target SCO4117 sequence | This study |
| pCRISPR-SCO4117A | pCRISPR-SgSCO4117 harboring a 2.5 kb fragment used to create the SCO4117 knockout | This study |
| pCRISPR-SCO4117B | pCRISPR-SgSCO4117 harboring a 2.2 kb fragment used to create the truncated SCO4117 gene harboring the sigma factor domain | This study |
| PRIMERS | ||
| SCO134 | GACGTGCTGCTGGTCATAGC | This study |
| SCO4117R | GGGACTAGTGTACGCCGCCGAAGTGG | This study |
| SCO159 | GGAGGCGATGTCCATCTGTT | This study |
| SCO160 | TCAACGCTCATCGCGGAAAG | This study |
| SCO163 | CTGCAGGCCCAGTCCCGCTTCAC | This study |
| SCO164 | GATATCACTCCTGGTCCTCGACAACT | This study |
| SCO4848F | CGTCGTATCCCCTCGGTTG | Gonzalez-Quiñonez et al., 2016 |
| pMS82R | GAGCCGGGAAAGCTCATTCA | Gonzalez-Quiñonez et al., 2016 |
| SCO197 | CATGCCATGGTTCCGCGATGAGCGTTGAAAGTTTTAGAGCTAGAAATAGC | This study |
| sgRNA-R | ACGCCTACGTAAAAAAAGCACCGACTCGGTGCC | Tong et al., 2015 |
| SCO4117-FA | GGGGGATATCACGACCGTGATCTCGGCC | This study |
| RT-SCO4117-F | CGACGACACGGCCTACGA | This study |
| RT-SCO4117-R | CGCACCGCCTGAAGCAT | This study |
| hrdB-F | CGCGGCATGCTCTTCCT | Kurt et al., 2013 |
| hrdB-R | AGGTGGCGTACGTGGAGAAC | Kurt et al., 2013 |
| bldN-F | CTCACCAGCGAGACCTTTCTGC | This study |
| bldN-R | TCGTTGGCGTCGAGCATCT | This study |
| SCO1178-F | TCAAGGTCCGGCAGGTCTA | This study |
| SCO1178-R | CCGTCCTCCTGCTTGGT | This study |
| chpA-F | CTCGTCCTCGTCCTCGACTT | Straight et al., 2006 |
| chpA-R | GTCGTTCTCGCACTTGTTGC | Straight et al., 2006 |
| chpH-F | CACCGGTGGTCTGGTTCTC | Straight et al., 2006 |
| chpH-R | ATCACGGAGATCGTGTTGC | Straight et al., 2006 |
| SCO2748-F | GAGATCACCCCGAAACTGG | This study |
| SCO2748-R | AAGTGCCAGTCGATGACGTT | This study |
| actVA2-F | ACTACGCCTCCCAGAACCTC | This study |
| actVA2-R | TTGTGCCCGCCGATGTC | This study |
| redF-F | CGGAGAACAAGGGCAAGC | This study |
| redF-R | CAGGGGGATGGCGAAG | This study |
| actII4-F | GCGGCTTTTTGGAATGC | This study |
| actII4-R | GCAGGGTCTCGTTCAGC | This study |
| SCO215 | CATGCCATGGCAGCTCGCTCCACTGGTAGAGTTTTAGAGCTAGAAATAGC | This study |
| SCO216 | GATATCCGGGAACCAACGTCGCACGG | This study |
| SCO217 | CGGTTCCGCAGGCTCCTACGCGGAAAGCCCCCGCCAAC | This study |
| SCO218 | GTTGGCGGGGGCTTTCCGCGTAGGAGCCTGCGGAACCG | This study |
| SCO219 | GATATCTGCAGGTGCGGATGATGCAC | This study |
| SCO220 | GATATCCGTATCTCCTTCCCCGAACG | This study |
| SCO221 | CGGTTCCGCAGGCTCCTACTGGAGGTAGGCCTGCTTG | This study |
| SCO222 | CAAGCAGGCCTACCTCCAGTAGGAGCCTGCGGAACCG | This study |
| Mut4117F | CCCTGTCGCAACCTCTGC | This study |
| Mut4117R | GACGGGCACCCTGCG | This study |