TABLE 3.
Oligonucleotides used in this study
| Name | Sequence | |
|---|---|---|
| Construction of pK19mobsacB-glnX2 and pK19mobsacB-glnX-glnH-pknG | ||
| ΔglnX_1 | TATATAGTCGACGCGTTCTGGATCGGCGGGAAAAGG | |
| ΔglnX_2 | CCCATCCACTAAACTTAAACAAGAAGTATCCTTCTCCGTTAACGG | |
| ΔglnX_3 | TGTTTAAGTTTAGTGGATGGGTCGATTCAGGGCAGTGGCACTGGT | |
| ΔglnX_4 | TATATAGGATCCAAAGCAGTCAAGGTTTCTGCCGGT | |
| ΔpknG_3 | TGTTTAAGTTTAGTGGATGGGGCGAATGCCGTGCGG | |
| ΔpknG_4 | GATTCTAGAGTTAGCTATCGCTAGGTACG | |
| Confirmation of gene deletion by colony-PCR | ||
| ΔglnX_fw | ATTCATCAAGGTAGCCAATACTTTC | |
| ΔglnX_rv | GCTCCTCAGGGGTCAGATCAT | |
| ΔpknG_rv | GATCGATCCAGAGCGTAACGC | |
| Construction of pJC1-glnXProm | ||
| glnXProm-fw | TATAGGATCCGCGTTCTGGATCGGCGGGAAAAG | |
| glnXProm-rev | TATAGTCGACTTATAAGTACTCCTGCAAACGGGGG | |
| Construction of pET-TEV-odhI-R87A and pET-TEV-odhI-R87P by site-directed mutagenesis | ||
| odhI_R87A fw | CTTGATGATGTCACCGTTTCAGCTCGCCACGCAG | |
| odhI_R87A rv | CTGCGTGGCGAGCTGAAACGGTGACATCATCAAG | |
| odhI_R87P fw | TGTCACCGTTTCACCTCGCCACGCAGAG | |
| odhI_R87P rv | ACAGTGGCAAAGTGGAGCGGTGCGTCTC | |
| Construction of pET-TEV-glnHΔSP | ||
| glnHΔSP fw | TATACATATGACTCCAACACCTGTGGAACCG | |
| glnHΔSP rv | TATACTCGAGTTATCCTTCATCGTTTTCTGT | |
| Construction of pET-TEV-glnHcore | ||
| glnHcore fw | CCTGTATTTTCAGGGCCATATGCCACTGCCACCGGATTCTTC | |
| glnHcore rv | TGGTGGTGGTGGTGGTGCTCGAGTTATGGCATGTACTGCAGCTGTG | |
| Construction of pET-TEV-glnHcore-S163T-T165S | ||
| glnH_S163T-T165S fw | ATGTAGATATTGTGATTCGTACGGTCTCCATCACCGACGAACGCGCC | |
| glnH_S163T-T165S rv | GGCGCGTTCGTCGGTGATGGAGACCGTACGAATCACAATATCTACAT | |
| Construction of pAN6-glnH | ||
| glnH fw | GACAGTCATATGCACGCTTTTCGACGC | |
| glnH rv | GACGCTAGCTCCTTCATCGTTTTCTGTC | |
| Construction of pAN6-glnH-C27A by site-directed mutagenesis | ||
| glnH_C27A fw | AACGCTGCTTGCTTCCGCCACTCCAACACCTGTG | |
| glnH_C27A rv | CACAGGTGTTGGAGTGGCGGAAGCAAGCAGCGTT | |
| Construction of pPREx2-glnX-(F1 to F7)-phoA and pPREx2-glnX-(F1 to F7)-lacZ | ||
| glnX fw | TGCAGAAGGAGATATACATATGATCCGGGATGGAAATG | |
| glnX-F1 rv | CGTCTGGCCTCTCGAGGCAGGCTGGCCGAAAGTTTCCTG | |
| glnX-F2 rv | CGTCTGGCCTCTCGAGGCAACAGGCTCCGCATTAGTG | |
| glnX-F3 rv | CGTCTGGCCTCTCGAGGCGTTGTAAAGCTCAGACGCCATC | |
| glnX-F4 rv | CGTCTGGCCTCTCGAGGCCCTGCGCGTAATCCGCATCAAC | |
| glnX-F5 rv | CGTCTGGCCTCTCGAGGCATTCAACGGCCCCGACGCTTC | |
| glnX-F6 rv | CGTCTGGCCTCTCGAGGCGCGAGAATCCGCGATCAG | |
| glnX-F7 rv | CGTCTGGCCTCTCGAGGCTAAGTACTCCTGCAAACG | |
| lacZ fw | GCCTCGAGAGGCCAGACGGGCCAGGCCTTGGGCCCTACCATGATTACGGATTCAC | |
| lacZ rv | GTAAAACGACGGCCAGTGTTATTTTTGACACCAGACCAAC | |
| phoA fw | GCCTCGAGAGGCCAGACG | |
| phoA rv | GTAAAACGACGGCCAGTGTTATTTCAGCCCCAGGGCGGCTTTC | |
| Amplification and sequencing primer for odhI, odhA, and pknG | ||
| odhI-78b-up | CAGGAAATTCTAGGATCTTACGGA | |
| odhI-76b-dwn | GGCATTCTATACACAAAACGGTTG | |
| odhA-up80b-fw | AGCCAACGACCAACGTTACAG | |
| odhA-988b-rv | TGCGCAGGAATTCACCGGA | |
| odhA-862b-fw | TGTCGGTTCCATGGATTACC | |
| odhA-2051b-rv | GTGGTGAAGCCGATCTGGTT | |
| odhA-1982-fw | CCAGAAACCATCAACCTGGC | |
| odhA-dwn75b-rv | GCAGCAAAAAGGGCCGTATGCTGTGT | |
| pknG-up150b-fw | GGTTGATTCGGCAGGTAAACTAC | |
| pknG-1300b-rv | GTTTGCCGTCGCGGACTGCC | |
| pknG-1052b-fw | CGATATTTTCACCATCGGACGC | |
| pknG-dwn150b-rv | CGCAGAAATCACATCAGCCCCAAT |