TABLE 1.
Oligonucleotides used for mutagenesis
| Target residue(s) | Sequence (5′-)a | Substitution(s) |
|---|---|---|
| XylA N65 | GGCGCAACACTGGTGACTTTGTCGGTGG | D65 |
| XylA Q144 | GTCCACCCGTTACAACCAGCCTTCGATC | N144 |
| XylA V151 | CTTCGATCGACGGTACTCAGACCTTCAACC | T151 |
| XylB N62 | GGCAACCGTGGTGACCACGTCGGTGG | D62 |
| XylB library | ||
| T38P39S40A41 | GTTGCCGTTGGTGTAGGTGAC(A/C)(G/T)CGC(T/C)G(G/C)(C/G)G(G/C)(T/C)ATCGGTCCAGAAGGAGAAG | T38, G38, S38, A38; P39, G39, R39, A39; S40, G40; A41, D41, E41 |
| N62H63 | CCTTACCACCGACG(T/A)(G/A)GT(T/C)ACCAC GGTTGCCGTTC | N62, D62; H63, F63, L63, Y63 |
| C141-D148C149 | CTGCTGGAAGGTCTTG(C/G)(A/T)ACCCTCGATGGAAGGAGCA(C/T)(A/T)GTTGCGAGTGCTG | C141, N141, Y141, S141; C149, T149, S149 |
a
Degeneracies are indicated by parentheses, underlined nucleotides correspond to mutated codons, and boldface nucleotides correspond to mutations.