Evidence that SOS induction in the ?mreB, ?mreC, and ?mreD cells was not a secondary effect of their spheroidal shape came from study of a pbp2ts strain which, when grown under
nonpermissive conditions, also undergoes a rod-to-sphere transition.
Confluent podocytes grown on 6-well plates originating from both permissive and nonpermissive conditions were treated for 1 hour with heme in HBSS (Hank's Balanced Salt Solution).
At ~50-60% confluence, the cells were transferred to 37[degrees]C (nonpermissive condition) for 14 days to silence the temperature sensitive tsA58 T antigen.
The earliest literature has offered support and a rationale for providing counseling services under
nonpermissive conditions (Arbuckle & Boy, 1961; Beier, 1952; Gometz & Parker, 1968; Snoxell, 1960).
The mutants are pooled, and the culture is split and grown under permissive and nonpermissive conditions. For essential genes, there is no permissive condition.
The initial set of random insertion mutants is arrayed on a master and then pooled and grown under permissive and nonpermissive conditions, as above.
