I have made a number of ts mutations in my favorite gene (I don't know what it
does, I only know it's essential). I did the usual plasmid shuffle and defined
my permissive temperature as 30¼ C. Now I have mutants that don't grow at 37¼
C. and it turns out that most of them are also VERY unhappy at room temp too.
This is true on plates as well as liquid. I wasn't expecting this. Even at the
permissive temp the cells don't look happy under the microscope (big, swollen
cells), but they definitely grow (about 2 hour doubling time). Meanwhile, our
WT is very happy at all three temps.
Has anyone seen this or have any words of inspiration?
bawagan at mbcl.rutgers.edu