TABLE 1.
Oligonucleotide sequences used for site-directed mutagenesis of colH
| Enzymea | Sequenceb |
|---|---|
| Wild type | 1648GAAGAATTATTTAGACATGAATATACACATTATTTGCAAGGAAG1691 |
| H415F | 1648GAAGAATTATTTAGATTTGAATATACACATTATTTG |
| H415A | 1648GAAGAATTATTTAGAGCTGAATATACACATTATTTG |
| E416D | 1651GAATTATTTAGACATGATTATACACATTATTTGC |
| E416Q | 1651GAATTATTTAGACATCAATATACACATTATTTGC |
| H419R | 1661GACATGAATATACACGTTATTTGCAAGGAAG |
| Wild type | 1723AAACTTTATGACAATGATAGATTAACTTGGTATGAAGAAGGTGGAGCAGAATTATTTGCAGGTTCTACT1791 |
| N439A | 1723AAACTTTATGACGCTGATAGATTAACTTGGTATGAA |
| E446Q | 1741AGATTAACTTGGTATCAAGAAGGTGGAGCAGAATTA |
| E446A | 1741AGATTAACTTGGTATGCAGAAGGTGGAGCAGAATTA |
| E446D | 1741AGATTAACTTGGTATGACGAAGGTGGAGCAGAATTA |
| E447Q | 1744TTAACTTGGTATGAACAAGGTGGAGCAGAATTA |
| E447A | 1744TTAACTTGGTATGAAGCAGGTGGAGCAGAATTATTT |
| E447D | 1744TTAACTTGGTATGAAGACGGTGGAGCAGAATTATTT |
| E451Q | 1756GAAGAAGGTGGAGCACAATTATTTGCAGGTTCTACT |
| E451A | 1756GAAGAAGGTGGAGCAGCATTATTTGCAGGTTCTACT |
| E451D | 1756GAAGAAGGTGGAGCAGACTTATTTGCAGGTTCTACT |
a
Mutant enzymes are designated by the amino acid residue replaced, its position, and the amino acid that was substituted for it.
b
Nucleotide positions are numbered as in reference 35. Nucleotide substitutions are underlined.