TABLE 3.
Effects of promoter mutations on rsmY promoter activity
| Construct | Sequencea |
lacZ expression level (103 Miller units) (mean ± SD)b |
|||
|---|---|---|---|---|---|
| Wild type | gacA mutant | rsmA rsmE mutant | rsmX rsmY rsmZ mutant | ||
| Wild type (pME7699) | TGTAAGTCATCTCTTACATAACGTGCTGCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 15.5 ± 0.9c | 0.9 ± 0.1 | 2.3 ± 0.1 | 24.0 ± 1.1 |
| UAS Δ6 (pME7695) | ------TCATCTCTTACATAACGTGCTGCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 0.5 ± 0.0 | 0.5 ± 0.0 | 0.6 ± 0.0 | 0.4 ± 0.0 |
| UAS mut12 (pME7689) | aagtctTCATCTcaatgtTAACGTGCTGCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 0.6 ± 0.0 | 0.3 ± 0.0 | 0.9 ± 0.0 | 0.4 ± 0.0 |
| −10 box mut3 (pME7696) | TGTAAGTCATCTCTTACATAACGTGCTGCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAtaaTACTTCACA | 15.5 ± 1.1 | 1.0 ± 0.0 | 2.8 ± 0.2 | 26.9 ± 1.3 |
| Linker mut6 (pME7682) | TGTAAGTCATCTCTTACATAAtagcCaaCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 2.0 ± 0.2 | 0.3 ± 0.0 | 0.5 ± 0.0 | 5.0 ± 0.1 |
| Linker mut21 (pME7694) | TGTAAGTCATCTCTTACAgtgatcagacagctggtgaatTATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 0.3 ± 0.0 | 0.2 ± 0.0 | 0.5 ± 0.0 | 0.3 ± 0.0 |
| Linker mut44 (pME7684) | TGTAAGTCATCTCTTACAcgcgctgtaattcaatgcgccttctgcgcaccgggctttgtgttTAATCTACTTCACA | 3.5 ± 0.2 | 0.9 ± 0.1 | 0.9 ± 0.1 | 4.5 ± 0.0 |
| Linker Δ3 (pME7683) | TGTAAGTCATCTCTTACA---CGTGCTGCTGATCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 |
| Linker Δ3bis (pME7685) | TGTAAGTCATCTCTTACATAACGTGCTGC---TCGTCCATATCGGTAGATCTCGGCTGAAGCTAATCTACTTCACA | 0.4 ± 0.0 | 0.4 ± 0.0 | 0.4 ± 0.0 | 0.4 ± 0.0 |
a
The UAS, artificially introduced BglII sites, and the −10 hexamer are shown in italics, with underlining, and in bold, respectively. Lowercase letters indicate mutational changes.
b
Values were determined at an OD600 of 2.0 to 2.5 for strains CHA0 (wild type), CHA89 (gacA mutant), CHA1009 (rsmA rsmE mutant), and CHA1144 (rsmX rsmY rsmZ mutant) grown in NYB.
c
The same value was found with pME6916 (rsmY-lacZ), which does not contain the artificial BglII site.