Gal Induction

Michael Forte forte at ohsu.edu
Fri Apr 4 13:31:21 EST 1997


Hi,

I need some expert advice on some problems we seem to be having with Gal
induction of specific constructs in yeast.  Basically, we are trying to
inducibly express a GST fusion of our protein of interest.  The construct
is in pEGKT and transformed into the Gal+ strain YM1372.  Cells are grown
overnight in raffinose and then transfered into Gal media (glucose-free
Gal) either in the presence or absence of raffinose.  We then take time
points (2-24 hrs) and look for GST expression on Westerns.  First, we are
clearly getting some expression even in the absence of Gal induction. 
Second, in some constructs, there appears to be no induction on Gal
addition; expression is unchanged from the uninduced levels.  I understand
that there is going to be some protein specific variation but is this
typical or are there tricks to getting this to work better?  Any tips
greatly appreicated.

Mike Forte



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