Titering Gal1 promoter

Mark Walberg walberg at simmons.swmed.edu
Thu Aug 31 12:11:23 EST 1995


Regarding titering the GAL1 promoter with mixtures of glucose and 
galactose:  I think that this is going to be very difficult to do in a 
controlled way.  In such mixtures I would expect a diauxic growth 
pattern with consumption of glucose first, then as glucose concentration 
falls the GAL1 promoter will become activated. 
    If you really want to test a range of expression levels, it would be 
better to use a gratuitous inducer.  Didier Picard's lab has made just 
such a system for the GAL1 promoter.  They made a construct that is a 
fusion of GAL4 DNA binding domain/transactivation domain/estrogen 
binding domain of ER.  The presence of this protein allows control of 
the level of activation of the GAL1 promoter by changing the estrogen 
concentration-even in glucose containing media.  The basal transcription 
rate without estrogen might be a little higher than normal for GAL1 
though-I can't remeber if it is significant or not.  The paper is in 
GENE about two years ago.  I don't have the full reference here.
walberg at simmons.swmed.edu





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