Table 2.
Primers and plasmid templates used to construct ADPRibase-Mn mutants
| Mutations | Forward mutagenic primersa | Template | |
|---|---|---|---|
| 1 | F37A + L196F + C253G | CCGGACGCGTCTGACAATGTGGGCCTGGCCTGGAACTACAG | F37A + L196F-wtb |
| 2 | F37A + L196F + D250A + C253G | CCCATTTACCCGGACGCGTCTGCAAATGTGGGCCTGGCCTGGAAC | 1 |
| 3 | F37A + L196F + V252A + C253G | CCCGGACGCGTCTGACAATGCTGGCCTGGCCTGGAACTACAGAG | 1 |
| 4 | F37A + L196F + V252G + C253G | CCCGGACGCGTCTGACAATGGTGGCCTGGCCTGGAACTACAGAG | 1 |
| 5 | F37A + L196F + V252A + C253G + T279A | GGATCCTCAGAGTAGCCACCATCATGTGCGTGACCAGC | 3 |
| 6 | C253V | CGGACGCCTCTGACAATGTGGTCCTGGCTTGGAACTACAG | Wild-typeb |
aReverse mutagenic primers were the exact reverse complement of those shown in the table
bTemplate described elsewhere55.