TABLE 3.
BacTH analysis of VraG-GraS and select mutant interactions
| Hybrid protein pair | β-Galactosidase activity (U/mg)a |
|---|---|
| GraS + VraG | 89.48 ± 3.61 |
| ΔEL::DYDFPIDSL GraS + VraG | — |
| ΔEL::AYEISVESV GraS + VraG | — |
| ΔEL::Ala9 GraS + VraG | — |
| GraS + ΔEL VraG | — |
| GraS + ΔGL VraG | 41.60 ± 1.87 |
| GraS + ΔGL::Gly10 VraG | — |
Complementation between hybrid proteins was quantified according to the BacTH quantification described in Materials and Methods. Each interaction represents the average of three biological replicates with standard deviation values. pUT18C-Zip plus PKT25-Zip represented the positive control for this assay and produced an average of approximately 500 U of β-galactosidase activity. Empty pKT25 and pUT18 vectors were used as negative controls and generated less than 5 U of β-galactosidase activity on average. One unit of β-galactosidase activity corresponds to 1 nmol of ONPG hydrolyzed per min at 28°C per mg of bacterial dry weight. Dashes indicate β-galactosidase activity of 5 U or less.