E.coli and toxic proteins

gc genecutl at mendel.berkeley.edu
Tue Jul 2 16:29:49 EST 1996


I am failing at my attempts to express a recombinant protein 
in E.coli.  The vector has a T7 promoter as well as a lac operator
and repressor, and I have tried using BL21 and BL21/pLysS for the
expression.  The problem is that the cells won't grow once the plasmid
is in them.  They will grow on a plate, actually, but once I put them
in liquid culture they don't grow at all.  I have tried growing a lawn
of cells on a plate, then scraping that off and resuspending them in
LB, then inducing, but I got no protein production.
Does anyone know any tricks to get a toxic protein expressed in E.coli?

--gc

...doom is like the handle of a pot, |
it's there, know it, have ice in your|
tea, marry, have children, visit your|   genecutl at mendel.berkeley.edu
dentist, do not scream at night even |
if you feel like screaming...   -cb  |



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