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Induction of Gal1 promoter on yeast lifts

Martin Hirst hirst at interomex.com
Wed May 29 12:07:40 EST 2002




Hi Mark,

You may be able to get away with plating on Rafinose.  Once the colonies
have formed, lift onto raf/gal (2%) plates for 4hrs to induce cDNA
expression.  Why do you think the cDNA's will be toxic?

Cheers

Martin





-------------------------------------------
 
Martin Hirst Ph.D


Interomex Biopharmaceuticals Inc.
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> I searched the archives, but didn't find the answer.
> 
> I have a library in pYES2, for induction with galactose.  It would be ideal
> to turn on expression of the cDNA on filter lifts, which would then be used
> to assay for the desired phenotype.  I am aware of this being done in liquid
> culture, but does anyone have or know of a protocol to do this on filters?
> 
> Our assumption is that the protein expressed will be toxic for yeast,
> therefore I don't believe that we could recover the colonies after induction
> by plating on Gal media.
> 
> If this is a trivial question, please forgive my ignorance.
> 
> --Mark
> 
> Mark T. Worthington
> 
> 

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