TABLE 1.
Locus (gene) | Direction of primera | Primerb | Sequence (5′→3′) |
---|---|---|---|
HP1512 (frpB4) | F | HP1512.1 | GTTTGTCTCTTGTATTGTCG |
R | HP1512.2 | CCACATATTGCTTCTTAAAG | |
HP1338 (nikR) | F | NikR-1 | ACTAGTCCATGGATACACCCAATAAAGAC |
R | N4_NotI | GCGGCCGCCGCTCAACAGTACCCTATCTATCCC | |
HP1511-1512 intergenic region | F | IG1511-12-1 | TCACGAACAGTGCTTGATTCAG |
R | HP1511-12-2 | CGATACGAAATTGATTTCTAAGC | |
HP1512 (frpB4) | F | HP1512RTF | TCTAGCTGCAACGACTGGGAATGT |
R | HP1512RTR | AGCGCGAGAGCCAGGTTAGGATA | |
HP1400 (fecA3) | F | HP1400RTF | TTGAGCGATCGCATTGAAGCTTGG |
R | HP1400RTR | AAGCCAATGTTGGTTGAGGGCATC | |
HP1010 (ppk) | F | HP1010RTF | GCGCGTTAGTCGTTTATGGCGTTT |
R | HP1010RTR | AGCGCTCAAAGGGTTGTAATTGCC | |
HP1077 (nixA) | F | HP1077RTF | CGTTTGATGCGGATCACATCGCTT |
R | HP1077RTR | TCTTCTAGCATCGGCGTGTGTTCT | |
HP1338 (nikR) | F | HP1338RTF | CGCCGTGCTTGTGGTGATTTATGA |
R | HP1338RTR | CCATGTGAATGTGCGTGGTGCATA | |
HP0073 (ureA) | F | HP0073RTF | CTGAATTGATGCAAGAAGGGCGCA |
R | HP0073RTR | CCACTTCATGGATCATGCTTGCCA |
F, forward; R, reverse.
Primers HP1512.1 and HP1512.2 were used for construction of the HP1512::cat mutants. Primers NikR-1 and N4_NotI were used for expression of recombinant NikR in pET30a. Primers used for real-time PCR are designated with RTF (real-time forward) and RTR (real-time reverse) in the primer name. HP1010 (ppk) served as a housekeeping gene for the quantitative real-time PCR experiments.