TABLE 1.
phoD promoter mutations
| phoD-lacZa plasmid or strain | DNA sequenceb | Promoter β-galactosidase activityc |
|---|---|---|
| −35 −25 | ||
| pSE7/MH5801/ wild type | TTCACAGTCGTTTAACA | 1.00 |
| pSE271b/MH5802 | TTCACAGTCGTgTAACA | 0.60 |
| pSE272b/MH5803 | TTCACAGTCGTTgAACA | 0.16 |
| pSE273b/MH5804 | TTCACAGTCGTTTAACg | 0.58 |
| pSE274b/MH5805 | TTCACAGTCGTTTAgCA | 0.22 |
| pSE282b/MH5809 | TTCACAGTCGTTTgACA | 0.14 |
| pSE283b/MH5810 | TTCACAGTCGTTTAAgA | 0.11 |
| pSE287b/MH5811 | TTCACAGTCGTTTAAtA | 0.62 |
| pSE288b/MH5812 | TTCACAGTCGTTcAACA | 0.41 |
| pSE289b/MH5813 | TTCACAGTCGTcTAACA | 0.20 |
| pSE290b/MH5814 | TTCACgGTCGTTTAACA | 0.42 |
| pSE291b/MH5815 | TTCAtAGTCGTTTAACA | 0.88 |
| pSE292b/MH5816 | TTCgCAGTCGTTTAACA | 0.28 |
| pSE293b/MH5817 | TTtACAGTCGTTTAACA | 1.00 |
| pSE294b/MH5818 | TcCACAGTCGTTTAACA | 0.35 |
| pSE295b/MH5819 | cTCACAGTCGTTTAACA | 0.45 |
| pSE330b/MH5827 | TTCAgAGTCGTTTAACA | 0.20 |
| pSE331b/MH5828 | TTgACAGTCGTTTAACA | 0.64 |
| pSE342b/MH5838 | TgCACAGTCGTTTAACA | 0.08 |
| pSE343b/MH5839 | gTCACAGTCGTTTAACA | 0.38 |
| pSE345b/MH5840 | TTCgCAGTCGTTTAgCA | 0.00 |
| pSE296b/MH5820 | TTCAGAGTCGcTTAACA | 0.92 |
| pSE297b/MH5821 | TTCACAGTCaTTTAACA | 0.90 |
| pSE298b/MH5822 | TTCACAGTtGTTTAACA | 1.00 |
| pSE299b/MH5823 | TTCACAGcCGTTTAACA | 0.89 |
| pSE300b/MH5824 | TTCACAaTCGTTTAACA | 0.77 |
| pSE332b/MH5829 | TTCACAGTgGTTTAACA | 1.06 |
| pSE333b/MH5830 | TTCACAGgCGTTTAACA | 0.99 |
| pSE336b/MH5831 | TTCACAGTCGgTTAACA | 1.08 |
| −55 −45 | ||
| Wild type | TTACAATCAGTTCACA | 1.00 |
| pSE348B/MH5843 | TTACAATCAGTTCgCA | 0.29 |
| pSE346b/MH5841 | TTAgAATCAGTTCACA | 0.24 |
| pSE347b/MH5842 | TTgCAATCAGTTCACA | 0.30 |
| −35 −25 | ||
| pSE374b/MH5846 | TTCACAGTCgatcgatcgaGTTTTAACA | 0.03 |
| −45 −35 | ||
| pSE344b/MH5837 | TTCACACTgatcgatcgaTCTTCACA | 0.14 |
| −35 −25 | ||
| pSE373b/MH5845 | TTCACAGTCgatcgGTTTAACA | 0.00 |
| −45 −35 | ||
| pSE341b/MH5836 | TTCACACTgatcgTCTTCACA | 0.00 |
| pSE275b/MH5806 | 5-bp insertion gacag at −90 | 0.17 |
| pSE301b/MH5825 | 10-bp insertion gacaggacag at −90 | 0.52 |
| pSE375b/MH5847 | 3′-199AAAATTCTAGTACACTT-183 5′ to | 0.02 |
| 3′-199AAGTGTACTAGAATTTT-183 5′ | ||
| pSE276b/MH8507 | 5′ Binding region deletion | 0.03 |
| pSE367 | Core binding region deletion |
Transformation of the parent strain, JH642, with the plasmids shown in the table resulted in the strains with wild-type or mutant phoD-lacZ fusions in single copy at the amyE locus.
The phoD nucleotide sequences from −22 to −38, −43 to −58, and −33 to −48 are shown. The wild-type consensus repeats are underlined. The nucleotide substitutions and insertions are in bold and lower case.
Cells containing the various phoD-lacZ fusions were grown in LPDM where β-galactosidase activity was determined every hour for 12 h. The highest level of induction attained before repression was used to calculate the specific activity of each promoter. The table gives an average of three independent assays and expresses them as fractions of wild-type activity.