Table 1.
In vivo target specificity results.
| Transposition efficiency | Fold change | Specificity | Fold change | |
|---|---|---|---|---|
| EK/LP | 3.1E-4 | 1.0 | 0.044 | 1.0 |
| R104A | 1.2E-4 | 0.39 | 0.099 | 2.2 |
| Q118A | 2.4E-4 | 0.77 | 0.8 | 1.8 |
| K160A | 5.0 E-5 | 0.16 | 0.065 | 1.5 |
| K164A | 4.0 E-6 | 0.013 | 0.14 | 3.1 |
| R189C | 5.0E-5 | 0.16 | 0.16 | 3.7 |
| R210A | 1.0 E-5 | 0.032 | 0.36 | 8.0 |
| K212M | 4.2E-4 | 1.4 | 0.037 | 0.8 |
| H213A | 7.0 E-5 | 0.23 | 0.065 | 1.5 |
| K249A | 2.6E-4 | 0.84 | 0.046 | 1.0 |
| R250A | 2.6E-4 | 0.84 | 0.046 | 1.0 |
Transposition events are represented by the total colony count of the Tet-resistant colonies, and specific events are represented by blue colonies. These were compared with a no-protein control plated on Kan plates to generate transposition efficiency. All of the mutant Tnps resulted in at least a slight, but reproducible net loss of transposition activity except for K212M. The blue : total ratio is used to determine specificity. K164A, R189C and R210A led to significant increases in specificity (increase in the blue : total ratio), while leading to a decrease in transposition efficiency. K212M displayed a slight, but reproducible decrease in specificity (decrease in the blue : total ratio).