Table 2 The primers used in this study
From: Expression regulation of multiple key genes to improve l-threonine in Escherichia coli
Names | Sequence (5′-3′) | Purpose |
|---|---|---|
ParcA-U-FR1 | TGTTAATTTGCAGCATGCATCAGGCAGGTCAGGGACTTTTGATAGCGCACAGACAGATAAAAATTACA | The thrR-arcA fragments with the homologous arm of arcA |
ParcA-D-RF1 | TGTTACCAACTCGTCTTCAACGATAAGAATGTGCGGGGTCTGCATGGTTGTTACCTCGTTACCTTTGG | |
ParcA-U-FR11 | AACGCAATTACGTACTTTAGTCATGTTACGCCGATCATGTTAATTTGCAGCATGCATCA | |
ParcA-D-RF11 | GCCTTCCGCTTCGAAAATACTTTTCAACGTGTTGCGTGTTACCAACTCGTCTTCAACGA | |
Pf-ParcA-F | GTCCTAGGTATAATACTAGT TTCGATTTAGTTGGCAATTTGTTTTAGAGCTAGAAATAG | pTargetF-arcA |
pf-ParcA-R | CTATTTCTAGCTCTAAAACAAATTGCCAACTAAATCGAAACTAGTATTATACCTAGGAC | |
Pf-ParcA-V-F | TTCGATTTAGTTGGCAATTT | |
ParcA-V-R | CGGGTTGAACGGTTTGGTG | Identify and sequence TWF051, TWF069, TWF074 and TWF078 |
ParcA-S-F | CGGGTTGAACGGTTTGGTG | |
ParcA-S-R | CGGGTTGAACGGTTTGGTG | |
PcpxR-U-FR | AAGTCATGGATTAGCGACGTCTGATGACGTAATTTCTGCCTATAGCGCACAGACAGATAAAAATTACA | The thrR-cpxR fragments with the homologous arm of cpxR |
PcpxR-D-RF | GGAAGTCAGCTCTCGGTCATCATCAACTAACAGGATTTTATTCATGGTTGTTACCTCGTTACCTTTGG | |
PcpxR-U-FR11 | GCTGCAAACATGCGTCAGGGGGTGTAAAACAACGTAAAGTCATGGATTAGCGACGTCTG | |
PcpxR-D-RF11 | CGTTGAAGCCTTCCATCTCGAGCAGCTCCTTTAATAGGGAAGTCAGCTCTCGGTCATCA | |
Pf-PcpxR-F | GTCCTAGGTATAATACTAGTTGACGTAATTTCTGCCTCGGGTTTTAGAGCTAGAAATAG | pTargetF-cpxR |
pf-PcpxR-R | CTATTTCTAGCTCTAAAACCCGAGGCAGAAATTACGTCAACTAGTATTATACCTAGGAC | |
Pf-PcpxR-V-F | TGACGTAATTTCTGCCTCGG | |
PcpxR-V-R | GCCGCCACCACATTAA | Identify and sequence TWF052 and TWF070 |
PcpxR-S-F | AGCAGCGTGGCTTAATGAACT | |
PcpxR-S-R | CATAATGACAGGCGTCTGGTGT | |
PgadE-U-FR | ACAAGGATGTAAATAATGAAAAGGATGACATATTCGAAACGATAGCGCACAGACAGATAAAAATTACA | The thrR-gadE fragments with the homologous arm of gadE |
PgadE-D-RF | AAAGCCCTGTAAAAGAAAAGAATCTTTCGTCATGAGAAAAATCATGGTTGTTACCTCGTTACCTTTGG | |
PgadE-U-FR11 | ATAGGCGTTTACTATATTGAACAACGATTCGGACAAGGATGTAAATAATGAAAAGGATG | |
PgadE-D-RF11 | TTTTATCATTTCGTGATTATCTTTCAACTGCCAAAAGCCCTGTAAAAGAAAAGAATCTT | |
Pf-PgadE-F | GTCCTAGGTATAATACTAGTATTCGAAACGATAACGGCTAGTTTTAGAGCTAGAAATAG | pTargetF-gadE |
pf-PgadE-R | CTATTTCTAGCTCTAAAACTAGCCGTTATCGTTTCGAATACTAGTATTATACCTAGGAC | |
Pf-PgadE-V-F | ATTCGAAACGATAACGGCTA | |
PgadE-V-R | GTTCCTGCCAGCATTCG | Identify and sequence TWF053, TWF071, TWF075, TWF079 and TWF081 |
PgadE-S-F | TGATAACTTATTCTTGGGCAGTA | |
PgadE-S-R | TTCATCAAGGATATGATTGTG | |
PpykF-U-FR | ACCGGATTCGCTTTCCGGCAGTGCGCCCAGAAAGCAAGTTTATAGCGCACAGACAGATAAAAATTACA | The thrR-pykF fragments with the homologous arm of pykF |
PpykF-D-RF | TTCGGTTTTCGGTCCGATGGTGCAAACAATTTTGGTCTTTTTCATGGTTGTTACCTCGTTACCTTTGG | |
PpykF-U-FR11 | TGTCACCTATCCTTAGAGCGAGGCACCACCACTTTCGTAATACCGGATTCGCTTTCCGG | |
PpykF-D-RF11 | TCATGCCAGCGTCCAGCATTTTAGCTAACATCTCTTCAGATTCGGTTTTCGGTCCGATG | |
Pf-PpykF-F | GTCCTAGGTATAATACTAGTTCTTAGTCTTTAAGTTGAGAGTTTTAGAGCTAGAAATAG | pTargetF-pykF |
pf-PpykF-R | CTATTTCTAGCTCTAAAACTCTCAACTTAAAGACTAAGAACTAGTATTATACCTAGGAC | |
Pf-PpykF-V-F | TCTTAGTCTTTAAGTTGAGA | |
PpykF-V-R | ACTTCCATACCGATCAGACCAT | Identify and sequence TWF054, TWF072, TWF076, TWF080, TWF082 and TWF083 |
PpykF-S-F | TGACAACTTCGGCACCAGA | |
PpykF-S-R | GTCGTTACCGCCTTCCAGT | |
PfadR-U-FR | GCGTAGTTAGCCCTCTGGTATGATGAGTCCAACTTTGTTTATAGCGCACAGACAGATAAAAATTACA | The thrR-fadR fragments with the homologous arm of fadR |
PfadR-D-RF | GAAACCCGCCGGGCTTTGCGCCTTAATGACCATGGTTGTTACCTCGTTACCTTTGG | |
PfadR-U-FR11 | GCCTTGATCCCTTTTTCTTCTTTTTGTCTGCTATCAGCGTAGTTAGCCCTCTGGTATGA | |
PfadR-D-RF11 | AAGCGGTTATTCCAGATACTTTCAATAATGTACTCTTCCGCGAAACCCGCCGGGCTTTG | |
Pf-PfadR-F | GTCCTAGGTATAATACTAGTAACTTTGTTTTGCTGTGTTAGTTTTAGAGCTAGAAATAG | pTargetF-fadR |
pf-PfadR-R | CTATTTCTAGCTCTAAAACTAACACAGCAAAACAAAGTTACTAGTATTATACCTAGGAC | |
Pf-PfadR-V-F | AACTTTGTTTTGCTGTGTTA | |
PfadR-V-R | GCCAAATCTCGCCACTC | Identify and sequence TWF055, TWF073 and TWF077 |
PfadR-S-F | GAGATCTCCATGATGGTTTCCCTTA | |
PfadR-S-R | GAGATCTCCATGATGGTTTCCCTTA | |
PiclR-U-FR1 | TGAAAATGATTTCCACGATACAGAAAAAAGAGACTGTCATGAAACGCATTAGCACCACC | The thrR1 fragments with the homologous arm of iclR |
PiclR-D-RF1 | TTTTCTGCCGCGTTTCGCGGGAATGGGTGCGACCATGGTTGTTACCTCGTTACCTTTGG | |
PiclR-U-FR11 | GTTCAGTAACTATTGCATTAGCTAACAATAAAAATGAAAATGATTTCCACGATACAGAA | |
PiclR-D-RF11 | GACTGAACCTGTCCAGTCGCTGGTGCGGTGGCAACGGCGGGTTTTCTGCCGCGTTTCGC | |
Pf-PiclR-F | GTCCTAGGTATAATACTAGTACAGAAAAAAGAGACTGTCAGTTTTAGAGCTAGAAATAG | pTargetF-iclR1 |
pf-PiclR-R | CTATTTCTAGCTCTAAAACTGACAGTCTCTTTTTTCTGTACTAGTATTATACCTAGGAC | |
Pf-PiclR-V-F | ACAGAAAAAAGAGACTGTCA | |
pf-V-R | TTGTCAGCAAGATAGCCAGA | |
ThrL-V-F | ATGAAACGCATTAGCACC | Identify and sequence TWF056, TWF057, TWF058, TWF059, TWF060, TWF061, TWF062, TWF063, TWF064 and TWF065 |
PiclR-V-R | CACCTGTTCTTCGCTCAGTTGG | |
PiclR-S-F | GTAGGCGTTGTGGATAGCGG | |
PiclR-S-R | TTGCCCAATGTCCCAGTTCG | |
PiclR-U-FR2 | ATTAGCTAACAATAAAAATGAAAATGATTTCCATAGCGCACAGACAGATAAAAATTACA | The thrR2 fragments with the homologous arm of iclR |
PiclR-D-RF2 | ATGGGTGCGACCATGACAGTCTCTTTTTTCTGTATCGTAAAAAAAAAGCCCGCACTGTC | |
PiclR-U-FR22 | ACTCATCGGATCAGTTCAGTAACTATTGCATTAGCTAACAATAAAAATGAAAATGATTT | |
PiclR-D-RF22 | GCGGTGGCAACGGCGGGTTTTCTGCCGCGTTTCGCGGGAATGGGTGCGACCATGACAGT | |
Pf-PiclR-F2 | GTCCTAGGTATAATACTAGTAGTCTCTTTTTTCTGTATCGGTTTTAGAGCTAGAAATAG | pTargetF-iclR2 |
pf-PiclR-R2 | CTATTTCTAGCTCTAAAACCGATACAGAAAAAAGAGACTACTAGTATTATACCTAGGAC | |
Pf-PiclR-V-F2 | AGTCTCTTTTTTCTGTATCG | |
PiclR-U-FR3 | CACCACGCAACATGAGATTTGTTCAACATTAAAAAACGCCTTAGTAAGTATTTTTCAGC | PthrL- thrR fragments with the homologous arm of iclR |
PiclR-U-FR33 | ACCATACTGGCATAAACGCATCTGTGGTAAAAGCGACCACCACGCAACATGAGATTTGT | |
PiclR-D-RF4 | TTTCGCGGGAATGGGTGCGACCATTGAGATACCTCTTCTTATTTTTCTGAGCAAAAAAAAAGCCCGCACTGTC | The thrRs1 fragments with the homologous arm of iclR |
PiclR-D-RF44 | CCTGTCCAGTCGCTGGTGCGGTGGCAACGGCGGGTTTTCTGCCGCGTTTCGCGGGAATGGGTGC | |
PiclR-U-FR5 | ATTAGCTAACAATAAAAATGAAAATGATTTCCACCAAAACCATAAGGAGGATTTACATGAAACGCATTAGCACCACC | The thrRs2 fragments with the homologous arm of iclR |
PiclR-U-FR55 | ACATGAGATTTGTTCAACATTAACTCATCGGATCAGTTCAGTAACTATTGCATTAGCTAACAATAAAAATGAAAATGATTT | |
PiclR-U-FR6 | ATTAGCTAACAATAAAAATGAAAATGATTTCCGTAGCTGAGACTCTAAGGGAGGCGTCACATGAAACGCATTAGCACCACC | The thrRs3 fragments with the homologous arm of iclR |
PiclR-U-FR7 | ATTAGCTAACAATAAAAATGAAAATGATTTCCGCATACATTCCACGACCATAGGAACCAACCCATGAAACGCATTAGCACCACC | The thrRs4 fragments with the homologous arm of iclR |
PiclR-U-FR8 | ATTAGCTAACAATAAAAATGAAAATGATTTCCAAGCCCATTTTACCATCGAAGAGGGACAGAATGAAACGCATTAGCACCACC | The thrRs5 fragments with the homologous arm of iclR |
PiclR-U-FR9 | ATTAGCTAACAATAAAAATGAAAATGATTTCCTCCTTTTAGGGCTATCGCGGAGGAGTAATGAAACGCATTAGCACCACC | The thrRs6 fragments with the homologous arm of iclR |
cysH-aspC-F2 | TAGCCACCTAGGAGGACTCCATGTTTGAGAACATTACCGCCG | The RBSs7-aspC fragment |
cysH-aspC-R2 | CTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAGGCACATAAAAAAGCCCGC | |
cysH-aspC-F1 | ACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGTGAAACCAGTACGGAAATCCTGGCGTCGC | The promoter of cysH |
cysH-aspC-R1 | GGAGTCCTCCTAGGTGGCTATGCCTTGCCTGATGCGAC | |
cys-aspC-F | TTCTGGTGGCCGGAAGGCGAAGCGGCATGCATTTACGTTGACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCAT | The PcysH-RBSs7-aspC fragment with the homologous arm of lacI |
cys-aspC-R | TGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCG | |
Pf-lacI-F | GTCCTAGGTATAATACTAGTTACGATGTCGCAGAGTATGCGTTTTAGAGCTAGAAATAG | pTargetF-lacI |
pf-lacI-R | CTATTTCTAGCTCTAAAACGCATACTCTGCGACATCGTAACTAGTATTATACCTAGGAC | |
Pf-lacI-V-F | TACGATGTCGCAGAGTATGC | |
lacI-aspC-V-F | CCTACGCTGGAACAATGGCAAACACTGGCACA | Identify and sequence TWF066, TWF067 and TWF068 |
lacI-aspC-V-R | GCTTCCACAGCAATGGCATCCTGGTCATCC | |
lacI-S-F | AACAGATCGAAGAAGGGGTTGA | |
lacI-S-R | GACGGCGCGTGCAGGGCC | |
cysJ-aspC-F2 | CGCATAACAATTCGTATCATAAGGAGACCATATGTTTGAGAACATTACCGCCG | The PcysJ-RBSs8-aspC fragment with the homologous arm of lacI |
cysJ-aspC-F1 | ACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGTGAAACCAGTAGTTGCGCAAAATCGCTGATT | |
cysJ-aspC-R1 | ATGGTCTCCTTATGATACGAATTGTTATGCGGTAAGCAAAGCTGTTTCTGCGC | |
cysD-aspC-F2 | AAGGCCAAAGACACTAAGAATTATTTATATGTTTGAGAACATTACCGCCG | The PcysD-RBSs9-aspC fragment with the homologous arm of lacI |
cysD-aspC-F1 | CCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGTGAAACCAGTACGGTGCCTTAAGCACTTTTTG | |
cysD-aspC-R1 | ATAAATAATTCTTAGTGTCTTTGGCCTTATGTTTCGACTATAGGGAGCGTAAG | |
RT-16SrDNA-F | TTTAATTCGATGCAACGCGAAGAACC | Transcriptional level |
RT-16SrDNA-R | CGGACCGCTGGCAACAAAGGATAAG | |
RT-iclR-F | CAGGGTTTCGTGCGTCAGGTTGG | |
RT-iclR-R | CGATAATAATCGCTTCGTGATCGCTTTG | |
RT-aspC-F | GAGAACATTACCGCCGCTCCTGC | |
RT-aspC-R | CCCGTCTCATCTTTATAGACACCAATCCC | |
RT-fadR-F | GGAAGTGCTGGCTACCGCTAATGAAG | |
RT-fadR-R | CATCCCGTTAAGAATCAGACCGTAAATCG | |
RT-cpxR-F | ACAGCATCTGGGTCAGGTGGTTTCC | |
RT-cpxR-R | CGGCAGTTTACGACGCAGGTTGG | |
RT-gadE-F | AGGCAATAAACCCTTCAAG | |
RT-gadE-R | TCGGCATCTAATTTCTCCAG | |
RT-pykF-F | GCGTTTCCATTGCTCTGCC | |
RT-pykF-R | GCTTTCAGGTGCTCACGGATTTC | |
RT-arcA-F | AGTCCCGTGCTGAACTGCTGAAG | |
RT-arcA-R | ACCGTGAATGGTGGCGATG |