Induction followed by ”rescue” of cells
In order to follow death/survival of cells through time, we performed induction in liquid medium, followed by “rescue” at different time-points, by spreading aliquots on repressive medium, as we had done previously (Maroc and Fairhead 2019). As shown on figure 3, survival rate drops in the first hours of induction, and then stabilizes at values that are similar to survival on plates. Thus the cut decides the fate of cells, and stopping the induction does not «rescue» cells. The percentage of red colonies rises from zero to 100% in 24 hrs forC. glabrata and C. nivariensis . For C. bracarensis , there is already a large number of red colonies at T0 and the percentage of red colonies does not reach above 80%. These observations may be explained by promoter leakage before the induction, that would promote the appearance of cells resistant to cutting through both unfaithful NHEJ and plasmid rearrangement.