Bacterial Transformation and MP Induction

Mid-log-phase (OD₆₀₀=∼0.5–0.8) cells of the strain of interest grown in 2xYT (United States Biological) were transformed with the desired MP, and recovered for 45 min in Davis rich media11 (link) to suppress MP induction. All transformations were plated on 2xYT in 1.8% agar (United States Biological) containing 40 μg ml⁻¹ chloramphenicol (Sigma Aldrich), 10 μg ml⁻¹ fluconazole (TCI America), 10 μg ml⁻¹ amphotericin B (TCI America) and 25 mM glucose (United States Biological) and grown for 12–18 h in a 37-°C incubator. Colonies transformed with the appropriate MP were picked the following day and grown in Davis rich media containing 40 μg ml⁻¹ chloramphenicol, 10 μg ml⁻¹ fluconazole and 10 μg ml⁻¹ amphotericin B for 12–18 h. Following overnight growth of the MP-carrying strains, cultures were diluted 1,000-fold into fresh Davis rich media containing 40 μg ml⁻¹ chloramphenicol, 10 μg ml⁻¹ fluconazole and 10 μg ml⁻¹ amphotericin B. The remainder of each experiment is described in each of the following sections.