Table 1. MccJ25r isolates from random mutagenesis and selection.
| Amino acid substitution | Codon substitution | Number of independent isolates | Complementation of rpoCts | MBC* (mg/ml) |
|---|---|---|---|---|
| none | none | - | ++ | 0.01 |
| 424 Asn→Ser | AAC→AGC | 1** | + | 0.1 |
| 428 Thr→Ile | ACT→ATT | 2 | ++ | 0.5 |
| 430 His→Leu | CAC→CTC | 3 | + | 0.5 |
| 464 Asp→Gly | GAC→GCC | 1 | ++ | 0.05 |
| 469 His→Arg | CAC→CGC | 1 | ++ | 0.05 |
| 504 Gln→Arg | CAG→CGG | 1 | ++ | 0.5 |
| 733 Ser→Phe | TCT→TTC | 1 | + | 0.5 |
| 738 Arg→Leu | CGT→CTT | 1 | ++ | 0.05 |
| 776 Thr→Ile | ACC→ATC | 1 | ++ | 0.5 |
| 779 Ala→Thr | GCT→ACT | 1 | ++ | 1 |
| 780 Arg→Cys | CGT→TGT | 1 | ++ | 1 |
| 782 Gly→Ala | GGT→GCT | 1 | ++ | 1 |
| 785 Asp→Gly | GAT→GGT | 1 | ++ | 0.5 |
| 786 Thr→Ile | ACC→ATC | 1 | ++ | 0.5 |
| 789 Lys→Arg | AAA→AGA | 1 | ++ | 0.5 |
| 789 Lys→Gln | AAA→CAA | 1 | ++ | 1 |
| 869 Cys→Arg | TGT→CGT | 1 | + | 1 |
| 933 Arg→Cys | CGT→TGT | 1 | ++ | 1 |
| 1244 Gln→Leu | CAG→CTG | 1 | ++ | 0.5 |
*
Minimum bacteriocidal concentration; defined as the lowest concentration of MccJ25 that yields a viable cell count of ∼0 after incubation 2 h at 37°C (see Experimental Procedures).
**
Isolated as double mutant 354 Val→Ile; 424 Asn→Ser; complementation and MBC data are for a single mutant constructed using site-directed mutagenesis.