TABLE 2.
Primers used in this study
| Purpose and primer name | Sequence (5′ to 3′)a | Use |
|---|---|---|
| Allelic exchange mutagenesis | ||
| hemP_UP_FW | CGACTCTAGAGGATCTACCGAAGAAGAGATTCGG | Upstream region of hemP |
| hemP_UP_RV | GCCCAACACCTCTCAGTCGATAGCTTGAAGCAAAC | |
| hemP_DOWN_FW | TGAGAGGTGTTGGGCATTA | Downstream region of hemP |
| hemP_DOWN_RV | CGGTACCCGGGGATCGCAGATCAAGCAGTGGCT | |
| hmuR_UP_FW | TAAAACGACGGCCAGTGCCATTCAACGAACGCGGCGTGACC | Upstream region of hmuR |
| hmuR_UP_RV | CTTGAGCTTCAGGGAACGGGCTGCGCCC | |
| hmuR_DOWN_FW | CCCGTTCCCTGAAGCTCAAGCGCGAGCG | Downstream region of hmuR |
| hmuR_DOWN_RV | CAGCTATGACCATGATTACGCCGGATCGAGCACGTTGATC | |
| Complementation analysis | ||
| pBBR_hemP_FW | CCACCGCGGTGGCGGCCGCTTACGGTGCGCCTGATTCGC | Construction of pCP1 |
| pBBR_hemP_RV | GCTGGGTACCGGGCCCCCCCGCGAGCGAATGGAGGCGC | |
| pBBR_hmuR_FW | CCACCGCGGTGGCGGCCGCTCCGGGCGCAGCCCGTTCC | Construction of pCR1 |
| pBBR_hmuR_RV | GCTGGGTACCGGGCCCCCCCACGAACGCGTCGCGCAGC | |
| XhoI_paraBC | CCCCTCGAGCCAAAAAAACGGGTATGGAG | Construction of pNITara |
| paraBC_NheI | CCCGCTAGCTTTATTATGACAACTTGACG | |
| pNITara_hemP_EcoRI | GGGGAATTCATTCATTCCATCGTATCGACAGT | Construction of pBAD-hemP |
| pNITara_hemP_HindIII | GGGAAGCTTCGAATGGAGGCGCACAAATAA | |
| qRT-PCR | ||
| dnaA_F | GGCGTTCGACGATTTCAAGC | Transcription of dnaA |
| dnaA_R | CGAACGCGTAGAAGAATTCC | |
| hemP_F | CAAGGTCACAGCCATATCAG | Transcription of hemP |
| hemP_R | CTTCGTCAGGATCAGCTTGC | |
| hmuR_F | GTATCTGCAGTACGCACATG | Transcription of hmuR |
| hmuR_R | GATTGCCGATCGAGGTATAG | |
| bfd_F | GTTTCCGATCGGAAGATTCG | Transcription of bfd |
| bfd_R | GTACCGACTCCTCGCATTTG | |
| Protein purification | ||
| NdeI_hemP | GGGCATATGACCGACACCATGCGCCC | Construction of pEThemP |
| XhoI_hemP | GGGCTCGAGCTTCGTCAGGATCAGC | |
| NdeI_ahpD | GGGCATATGGAATTCATCGATTCGATTAAGG | Construction of pETahpD |
| XhoI_ahpD | GGGCTCGAGTTCGCCTTCGGCGG | |
| EMSA | ||
| pNIT_hmuR_pro_NheI | GGGGCTAGCGTCTCCCGAATTGATCGCGCG | Upstream region of hmuR |
| pNIT_hmuR_pro_HindIII | GGGAAGCTTAGGTGGTCTCCCATGGTTGA | |
| pNIT_5041 | CTGACACCCTCATCAGTGCCAA | Construction of probe |
| pNIT_5548 | GTGAGAAATCACCATGAGTGACG |
a
Restriction sites are underlined.