Table 1.
Oligonucleotides used in substitution mutagenesis for 10 residues of Bst DNA pol LF.
| Mutagenesis | Primer name | Sequence |
|---|---|---|
| G310A | F (5′-3′) | GTGGTGCACCCCGTGACGGCGAAAGTGCACACGATGTTCAATCAG |
| R (5′-3′) | CTGATTGAACATCGTGTGCACTTTCGCCGTCACGGGGTGCACCAC | |
| G310L | F (5′-3′) | GTGGTGCACCCCGTGACGCTCAAAGTGCACACGATGTTCAATCAG |
| R (5′-3′) | CTGATTGAACATCGTGTGCACTTTGAGCGTCACGGGGTGCACCAC | |
| D540A | F (5′-3′) | CGCCTGTTGCTGCAAGTGCATGCAGAACTGATTTTGGAGGCGCCGAAAGA |
| R (5′-3′) | CTCTTTCGGCGCCTCCAAAATCAGTTCTGCATGCACTTGCAGCAACAGGCG | |
| D540E | F (5′-3′) | CGCCTGTTGCTGCAAGTGCATGAAGAACTGATTTTGGAGGCGCCGAAAGA |
| R (5′-3′) | CTCTTTCGGCGCCTCCAAAATCAGTTCTTCATGCACTTGCAGCAACAGGCG | |
| R412A | F (5′-3′) | CGAAGAAGACGTGACAGCCAACATGCACGCCAAGCGAAGGCCGTCAATT |
| R (5′-3′) | AATTGACGGCCTTCGCTTGGCGTGCCATGTTGGCTGTCACGTCTTCTTCG | |
| R412E | F (5′-3′) | CGAAGAAGACGTGACAGCCAACATGGAACGCCAAGCGAAGGCCGTCAATT |
| R (5′-3′) | AATTGACGGCCTTCGCTTGGCGTTCCATGTTGGCTGTCACGTCTTCTTCG | |
| K416A | F (5′-3′) | GTGACAGCCAACATGCGCCGCCAAGCGGCAGCCGTCAATTTTGGCA |
| R (5′-3′) | TGCCAAAATTGACGGCTGCCGCTTGGCGGCGCATGTTGGCTGTCAC | |
| K416D | F (5′-3′) | GTGACAGCCAACATGCGCCGCCAAGCGGATGCCGTCAATTTTGGCA |
| R (5′-3′) | TGCCAAAATTGACGGCATCCGCTTGGCGGCGCATGTTGGCTGTCAC | |
| G310A-D540E | F (5′-3′) | GTGGTGCACCCCGTGACGGCGAAAGTGCACACGATGTTCAATCAG |
| R (5′-3′) | CTGATTGAACATCGTGTGCACTTTCGCCGTCACGGGGTGCACCAC | |
| G310L-D540E | F (5′-3′) | GTGGTGCACCCCGTGACGCTCAAAGTGCACACGATGTTCAATCAG |
| R (5′-3′) | CTGATTGAACATCGTGTGCACTTTGAGCGTCACGGGGTGCACCAC |
Underlined sequences indicate the mutant codons.