Research Article

Role of GntR Family Regulatory Gene SCO1678 in Gluconate Metabolism in Streptomyces coelicolor M145

Table 2

Primers used in this work.

Primer nameSequencePurpose/PCR product

SCO1678-fAATAAAAGCTTCGTGACTGAAGAAGAGCGAATo amplify SCO1678 with 3-kb flanks
SCO1678-rAATAATCTAGAAGGGAGTAATGAGGCTACGA
SCO1678_acc_fGACTTTTTGTTCCCGTGACATGCCCCGTACGCTGAGTGCATGGATATCTCTAGATACCGTo amplify aac(3)IV for SCO1678 replacement
SCO1678_acc_rTCCTCGACTGACGACGTCTCCCCCTGCCGCCCCGGCGCTCAAACAAAAGCTGGAGCTC
SCO1678NcoI-fAATTAACCATGGGCAGCACACCGGGCCGGGGGCTSCO1678 ORF for protein production
SCO1678HindIII-rAATTAAAAGCTTGGGCGCCAGGATGTCCAGCT
SCO1678gusXbaIAATAATCTAGACAGGGGACCGATGGTGGTCTPromoter region of SCO1678 to clone into pGUS
SCO1678gusKpnIAATAAGGTACCGCACTCAGCGTACGGGGCAT
SCO1678 cmpl-fAATAAGGATCCAGGGGACCGATGGTGGTCTTSCO1678 with 500 bp promoter
SCO1678 cmpl-rAATAATCTAGATTCCTCGACTGACGACGTCT
SCO1679Cy5-fAGCCAGTGGCGATAAGCTGTGATCGCGGGGCCGAGGCy5-labeled gntR-K intergeneric region
SCO1679Cy5-rAGCCAGTGGCGATAAGGTTGCTGCATCGCACTCTCG
SCO1680Cy5-fAGCCAGTGGCGATAAGGCAGCCGGACGAGGCGGGAGCy5-labeled gntP promoter region
SCO1680Cy5-rAGCCAGTGGCGATAAGGGTGGTTCCCTTGCGGTGAT
SCO1681Cy5-fAGCCAGTGGCGATAAGCTGGCTGGTGAAGGAGTACTCy5-labeled gntZ promoter region
SCO1681Cy5-rAGCCAGTGGCGATAAGCGTCGTACTCCTGTTCCTGC
SCO1263Cy5-fAGCCAGTGGCGATAAGCAGGGCCAGCAGACCGCCCACy5-labeled SCO1263 promoter
SCO1263Cy5-rAGCCAGTGGCGATAAGCGCTCCATCCCAGCGGACGC
RTSCO1678-fTGCGAGTGGAACGTCTACGART-PCR analysis of SCO1678
RTSCO1678-rGTCCTCGAACATCACGTCGT
RTSCO1679-fAACATCGCCAAGATGACGGCRT-PCR analysis of SCO1679
RTSCO1679-rGCCCGTTCGGTGATCTCCTC
RTSCO1680-fTGTTCTTCGAGGTCGGCATCRT-PCR analysis of SCO1680
RTSCO1680-rCTTGAGCAGCATCAGCACGA
RTSCO1681-fTCGACATCCTGGTCAACAACRT-PCR analysis of SCO1681
RTSCO1681-rGGTGCTGAACTCCTCGTCCT
RTSCO4991-fACCGGTCTGATCTTCGGCATRT-PCR analysis of SCO4991
RTSCO4991-rATCCAGGCGGAGAACACCCA
RTgntR-K_fACTCGACGAGGTGCATGGACRT-PCR of gntR-K intergeneric region
RTgntR-K_rGCCGTCATCTTGGCGATGTT
RTgntK-P_fTGATCGAGGACCGGATGTCGRT-PCR of gntK-P intergeneric region
RTgntK-P_rTGGTGATGACCTTGTCCAGC
RTgntP-Z_fACATGTCGACCACGCACGCRT-PCR of gntP-Z intergeneric region
RTgntP-Z_rACCGTCGGTCACGTCGAACA
RThrdB_fCGAGGACGAGGCGACCGAGGAGRT-PCR analysis of hrdB gene
RThrdB_rCAGCTTGTCCTCGGCGAACAGA