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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?



Martin Hirst Ph.D

Interomex Biopharmaceuticals Inc.
3590 West 41st Ave.
Vancouver, BC
V6N 3E6

direct line:(604) 267-8005
home:       (604) 943-5856
fax:        (604) 267-6411

email: hirst at interomex.com


> 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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