[Yeast] Slow galactose induction

Z.J. Zhang via yeast%40net.bio.net (by ZZhang from uwyo.edu)
Tue Sep 4 16:27:43 EST 2007


Kerry:

One possibility is the slow maturation of your CFP protein, rather than
slow induction.

Depending on the version of the CFP, and where the CFP is tagged, it
could take hour(s) to mature.

Zhaojie

Zhaojie Zhang, Ph.D.
Director, Microscopy Core Facility
Department of Zoology and Physiology
University of Wyoming
Laramie, WY 82071
TEL: 307-766-3038
FAX: 307-766-5625
zzhang from uwyo.edu
http://www.uwyo.edu/microscopy



-----Original Message-----
From: Kerry Geiler [mailto:kgeiler from gmail.com] 
Sent: Tuesday, September 04, 2007 11:55 AM
To: yeast from magpie.bio.indiana.edu
Subject: [Yeast] Slow galactose induction

Hi,

I am looking for suggestions as to why my galactose induction process
takes 6 hours.  I am expressing CFP under a gal promoter in media with
no glucose and 2% galactose.  I am getting fluorescence, but it takes
6 hours to see it.  Any ideas why induction could be taking so long?

I grow the cells up in media containing no glucose (2%raffinose)
overnight.  Then I induce 150uL of this culture with 2% galactose and
measure fluorescence every 15 minutes.  I do not see fluorescence
distinguishable from background until about 6 hours later.

Thanks very much for any suggestions!  Please reply to
kgeiler from gmail.com.
Thanks,
Kerry

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