TABLE 2.
Site-directed mutagenesis of dcuS in the region coding for the periplasmic domain of DcuS in plasmid pMW181
| Mutation(s) | Sequence (5′→3′)a |
|---|---|
| DcuS(A164S) | CAAATTGGCGTGGTGTCTATCGGCCTTGAGTTAAGCC |
| DcuS(F149K) | CGCAGGCTTTACGCGTAAAAACCCCCATCTACGATGAAAATC |
| DcuS(T101G) | GATCTGCTGTTTATTGTCGTTGGCGATATGCAAAGTCTTC |
| DcuS(F120M) | CGTATTGGTCAGCCAATGAAAGGTGATGAC |
| DcuS(I125S) | CCATTTAAAGGTGATGAC/TCGCTTAAAGCGCTGAATGGCG |
| DcuS(F120M I125S) | CCAATGAAAGGTGATGACTCGCTTAAAGCGCTGAATGGCG |
| DcuS(F141S) | CTATCAATCGCGGTTCTCTGGCGCAGGCTTTAC |
| DcuS(Q159V) | CTACGATGAAAATCATAAAGTAATTGGCGTGGTGGCGATC |
| DcuS(M103A) | TGTCGTTACCGATGCGCAAAGTCTTCGCTACTCG |
| DcuS(I125S) | CCATTTAAAGGTGATGACTCGCTTAAAGCGCTGAATGGCG |
| DcuS(R107A) | CCGATATGCAAAGTCTTGCCTACTCGCATCCTGAAG |
| DcuS(H110A) | CTTCGCTACTCGGCACCTGAAGCCCAGC |
| DcuS(R147A) | GGCGCAGGCTTTAGCAGTATTTACCCCC |
| DcuS(F149A) | GCAGGCTTTACGCGTAGCTACCCCCATCTACG |
| pdcuS-Nde2 | ATTTACTTCTCGCATATGAGTGATATG |
| pdcuS-Hind | GACCAGATAAAGCTTCAGCGACTG |
a
Oligonucleotides for mutagenesis are complementary (reverse) to the sequences shown; nucleotides resulting in amino acid exchanges in DcuS are underlined.