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[Yeast] RE: Slow galactose induction (Kerry Geiler)

Carlos Hassun via yeast%40net.bio.net (by multifaro from gmail.com)
Thu Sep 6 11:27:59 EST 2007


PLEASE, I NEED CONVERT GLYCERIN TO METANOL; IF ANYWHERE HAVE A WAY PLEASE
SEND INFORMATION.
REGARDS
C. I. Hassun

2007/9/5, Wang, Xin-Xiang <WangXX from medimmune.com>:
>
> Hello Kerry,
>
> I used to induce yeast for GFP expression, also using Gal promoter.  I
> noticed the trace amount of repressing medium (raffinose in your case or
> glucose in my case) was essential for GFP induction.  If I washed the trace
> amount of repressing medium away with induction medium, GFP was not induced
> in even a week!  But with the trace amount of glucose medium in, induction
> took only 15 hours (not test shorter time).
>
> Did you wash away the raffinose medium before induction or not? Good luck!
>
> Xinxiang
>
> ________________________________
>
> From: yeast-bounces from oat.bio.indiana.edu on behalf of
> yeast-request from oat.bio.indiana.edu
> Sent: Wed 9/5/2007 1:02 PM
> To: yeast from magpie.bio.indiana.edu
> Subject: Yeast Digest, Vol 28, Issue 2
>
>
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> Today's Topics
>
>    1. Slow galactose induction (Kerry Geiler)
>    2. RE: Slow galactose induction (Z.J. Zhang)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Tue, 4 Sep 2007 13:54:57 -0400
> From: "Kerry Geiler" <kgeiler from gmail.com>
> Subject: [Yeast] Slow galactose induction
> To: yeast from magpie.bio.indiana.edu
> Message-ID:
>         <3fa3f1ca0709041054o692505aek8f75bbd8b3d1d072 from mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> 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
>
>
>
> ------------------------------
>
> Message: 2
> Date: Tue, 4 Sep 2007 15:27:43 -0600
> From: "Z.J. Zhang" <ZZhang from uwyo.edu>
> Subject: RE: [Yeast] Slow galactose induction
> To: "Kerry Geiler" <kgeiler from gmail.com>, <yeast from magpie.bio.indiana.edu>
> Message-ID:
>         <C9C1AF307F12AF4087DEF87CB0E6A4AD01A3881F from TELEGRAPH5.uwyo.edu>
> Content-Type: text/plain;       charset="US-ASCII"
>
> 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
>
>
>
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