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.