TABLE 3.

Primers used in this study

EntryPrimer nameApplication or functionSequence (reference)
1TrpRS1 MK54Detection of the S. coelicolor trpRS1 (SCO3334) transcript via RT-PCR5′-CAG GAG GTG GAC ATA TGA CAC GGG TTT TCA G-3′ (22)
2TrpRS1 MK134Detection of the S. coelicolor trpRS1 (SCO3334) transcript via RT-PCR5′-CAC GTA CCC GGG ATC GGC GGA CAG C-3′ (22)
3TrpRS2 MK133Detection of the S. coelicolor trpRS2 (SCO4839) transcript via RT-PCR5′-GCG TCT CCG GGT CGT CCA GGT ACT G-3′ (22)
4TrpRS2 MK135Detection of the S. coelicolor trpRS2 (SCO4839) transcript via RT-PCR5′-CGC GAC CTC GCG GAC CGC TTC AAC CAG-3′ (22)
5SAV3417 RT FORDetection of the S. avermitilis SAV3417 transcript via RT-PCR5′-CCA CCT CGG CAA CTA CCT C-3′
6SAV3417 RT REVDetection of the S. avermitilis SAV3417 transcript via RT-PCR5′-TTG CCG GTG TAC TTC TCC TC-3′
7SAV4725 RT FORDetection of the S. avermitilis SAV4725 transcript via RT-PCR5′-ATC TGA CGC TGG GGA ACT AC-3′
8SAV4725 RT REVDetection of the S. avermitilis SAV4725 transcript via RT-PCR5′-CCG TAC CGC TGG TTG AAG-3′
9SGR2702 RT FORDetection of the S. griseus NBRC 13350 SGR2702 transcript and corresponding transcript from the indolmycin producer S. griseus ATCC 12648 via RT-PCR5′-ACG ACG CCT TCT ACA TGG TC-3′
10SGR2702 RT REVDetection of the S. griseus NBRC 13350 SGR2702 transcript and corresponding transcript from the indolmycin producer S. griseus ATCC 12648 via RT-PCR5′-GCC TGG TAG AGC AGG ATG TC-3′
11SGR3809 RT FOR 1Detection of the S. griseus NBRC 13350 SGR3809 transcript via RT-PCR5′-ACA CCC GCC GTC ACC AC-3′
12SGR3809 RT REV 1Detection of the S. griseus NBRC 13350 SGR3809 transcript via RT-PCR5′-GAG AGC ATG ATG TCC GGT TC-3′
13SGR3809 RT FOR 2Detection of the S. griseus ATCC 12648 SGR3809 homolog transcript via RT-PCR5′-GGC TGC TCC TGG ACT ATC TG-3′
14SGR3809 RT REV 2Detection of the S. griseus ATCC 12648 SGR3809 homolog transcript via RT-PCR5′-GTC GGG GTC GTA GGT GAT G-3′
15SCO4839 KO DET FORCloning of the S. coelicolor trpRS2 locus located 93 nucleotides upstream of the translational start site5′-CCC CGC CCT TGT TAC AC-3′
16SCO4839 KO DET REVCloning of the S. coelicolor trpRS2 locus located 96 nucleotides downstream of the translational stop site5′-CAA CCG GAG TGA TGT GCA G-3′
17SAV4725 FORCloning of the S. avermitilis SAV4725 ORF5′-CAT ATG AAG CGG ATC TTC AGC-3′a
18SAV4725 REVCloning of the S. avermitilis SAV4725 ORF5′-CTC GAG TTA CGC CTC CAG CAA CC-3′b
19SCO3334 FORCloning of the S. coelicolor trpRS1 ORF5′-CAT ATG ACA CGG GTT TTC AGT G-3′a
20SCO3334 REVCloning of the S. coelicolor trpRS1 ORF5′-CTC GAG CTA CCG GGC CGC GTT C-3′b
21SGR3809 FORCloning of the S. griseus NBRC 13350 SGR3809 ORF5′-CAT ATG ACC ACA CCC GCC G-3′a
22SGR3809 REV 2.0Cloning of the S. griseus NBRC 13350 SGR3809 ORF5′-CTC GAG CTA CCG GAA CGC GCC CAT C-3′b
23SCO4839 L9F FORSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of L9F mutation5′-GAA CGT CCC CGT GTG TTC TCC GGT ATC CAG C-3′
24SCO4839 L9F REVSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of L9F mutation5′-GCT GGA TAC CGG AGA ACA CAC GGG GAC-3′
25SCO4839 Q13K FORSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of Q13K mutation5′-GTG CTC TCC GGT ATC AAG CCG ACC TC-3′
26SCO4839 Q13K REVSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of Q13K mutation5′-GAG CCG GAG GTC GGC TTG ATA CCG GAG-3′
27SCO4839 H48Q FORSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of H48Q mutation5′-GGT CGT CGA CCT GCA GGC GAT CAC GG-3′
28SCO4839 H48Q REVSite-directed mutagenesis of the S. coelicolor trpRS2 locus, creation of H48Q mutation5′-CGG GAC CGT GAT CGC CTG CAG GTC G-3′
  • a The engineered NdeI site is underlined.

  • b The engineered XhoI site is underlined.