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Titering Gal1 promoter

Trueheart 71562.2253 at COMPUSERVE.COM
Thu Aug 31 11:42:37 EST 1995


>jjschm at ccmail.monsanto.com (Jon Schmuke) wrote:
>>Hi all,
>>
>>Does anyone know the best way to titer expression from the Gal1 promoter?
..
>I don't know what the best way is, but we have had excellent luck growing cells
in 2% 
>galactose with varying amounts of glucose (up to 1%).  With 2%/1% gal/glu in
YEP 
>medium, expression from the GAL1 promoter is reduced 100-fold, as assayed by
western 
>of the expressed protein.  This is in a strain that started out as S288C
background 
>but was backcrossed several times to a good Gal+ strain.  

>Tim Stearns
>stearns at leland.stanford.edu
_______________________________
Tim (and all):
When you add glucose to galactose, don't the cells use up the glucose first, and
hence fully induce themselves when they're finished?  Do you have to catch them
real early, before this has happened?

Josh Trueheart
Cadus




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