Supplementary Figure 11: Fluorescence screen for retention of selenocysteine incorporation in evolved populations using seleno-smURFP.

Internal controls are located below the streaks are (from left to right) ΔsmURFP, smURFP-C52, seleno-smURFP (U52) and smURFP-S52. (a) β_Δ populations deficient in selenocysteine incorporation were non-fluorescent. Both the non-selenocysteine dependent population (β_CC) and the two selenocysteine dependent populations (β_UC and β_UU) had almost complete retention of the trait. (b) T_Δ populations deficient in selenocysteine incorporation were non-fluorescent. The selenocysteine dependent T_UC and T_UU populations had almost complete retention of the trait, although no transformants could be obtained from T_UC3 and only a small number from T_UU3. In contrast, retention was highly variable across the three T_CC populations which were not selenocysteine dependent. Note, in this composite image the T_UU2 plate was imaged after an additional 24 hours of incubation at 37 °C due to very slow growth of the streaked cells using an exposure time of 0.5 seconds. The internal controls are consistent with the other plates and clear differences are observed between the streaked cells and the non-fluorescent smURFP-S52 control.