Table 2 PCR primers and the targets of amplified fragments
From: ComCED signal loop precisely regulates nlmC expression in Streptococcus mutans
Primer | Primer sequence (5′ → 3′) | Sequence targeted |
|---|---|---|
ComE-F | tcCCATGGTAATGATTTCTATTTTTGTATTGGAAG | comE for protein expression |
ComE-R | tgCTCGAGTCATTCATTTTGCTCTCCTTTGAT | |
PnlmA-F | AAATTAGCTGGTAATGATAGTT | nlmAB promoter (PnlmAB) (−163 to −43) |
PnlmA-R | GCAACCAACATCTTTAGTATAA | |
Nest-nlmC-F | AAATCCAGCCTTGTTTTATTCC | Upstream region of nlmC used in nest PCR for the amplification of nlmC regulating region (−444 to + 103) |
Nest-nlmC-R | GAGCAATACCTGCTGCACAA | |
PnlmC-F | AAAGTAATATTTTCCTTAGGAC | nlmC promoter (PnlmC) (−197 to + 51) |
PnlmC-R | AAGTGCTTCATTATCCATTA | |
PnlmC314-F | GTCTTAATTTCTTTAAAGTC | Upstream region of nlmC promoter (PnlmC314) (−314 to −166) |
PnlmC314-R | TACATTATGTGTCCTAAGGA | |
PnlmC419-F | GTTAGCCTATCTTATTTTCC | Upstream region of nlmC promoter (PnlmC419) (−419 to −295) |
PnlmC419-R | GACTTTAAAGAAATTAAGAC | |
Erm-H3 | gaAAGCTTCCGGGCCCAAAATTTGTTTGAT | erythromycin (erm) for pLacZ construction |
Erm-XO | aaCTCGAGTCGGCAGCGACTCATAGAAT | |
upComED-KN | GAACATAATTTACAGCGGTTCATA | comE and its upstream region (−231 to + 371) for comED mutant construction |
upComED-H3 | ggAAGCTTCAATGCGGTGGGAGAACT | |
downComED-XO | gaCTCGAGTTAGGCGGGCAATCATATTC | comD and its downstream region (+ 289 to + 1069) for comED mutant construction |
downComED-BH | AGCAGCCTCAATGGCATTAT | |
upComC-KN | ATCTGAACAAGCAGGGGAGA | comC and its upstream region (−752 to + 13) for comC mutant construction |
upComC-H3 | gcAAGCTTGTGTTTTTTTCATTTTATATCTCC | |
downComC-XO | taCTCGAGTCCGGCTGTTTAACAGAAGTT | comC and its downstream region (+ 98 to + 940) for comC mutant construction |
downComC-BH | GGCACAAAAGGAAGCTCAGA | |
upComX-KN | TTAGTTTGTAATGCGAGGTG | comX and its upstream region for comX mutant construction |
upComX-H3 | gcAAGCTTAACGACGTTTCTGACTTTCT | |
downComX-XO | tgCTCGAGGAAGTCGGTGAGATTGAACA | comX and its downstream region for comX mutant construction |
downComX-BH | GACCAAGATTGATTTTAGCC | |
LacZ-XF | gaTCTAGAGCCACACAGGAAACAGCTATGAC | Beta-Galactosidase (lacZ) used as a reporter in the analysis of promoter activity |
LacZ-SR | aaGTCGACGAAGTAGGCTCCCATGATAAA | |
LnlmA-BF | gcGGATCCGTATCGGAAGAATTATCTGG | nlmA upstream region (−741 to + 56) used in pA-LacZ construction |
LnlmA-XR | atTCTAGAGGTTGAAAGTGTTTGGCTGT | |
LnlmC-BF | aaGGATCCTTTTATCTTCTCATCCACGAC | nlmC upstream region (−523 to + 122) used in pC-LacZ construction |
LnlmC-XR | agTCTAGAGTAACCTTGCCCAGCACCTA |