Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Animal and Molecular Model Systems > Yeast Forum
Register Search Today's Posts Mark Forums Read

Yeast Forum Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


Slow galactose induction (Kerry Geiler)

Slow galactose induction (Kerry Geiler) - Yeast Forum

Slow galactose induction (Kerry Geiler) - Discuss research on yeast the model organism. Post questions and topics on the molecular biology and genetics of Saccharomyces cerevisiae and other yeast species.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 09-05-2007, 06:29 PM
Wang, Xin-Xiang
Guest
 
Posts: n/a
Default Slow galactose induction (Kerry Geiler)



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: [Only registered users see links. ] on behalf of [Only registered users see links. ]
Sent: Wed 9/5/2007 1:02 PM
To: [Only registered users see links. ]
Subject: Yeast Digest, Vol 28, Issue 2



Send Yeast mailing list submissions to
[Only registered users see links. ]

To subscribe or unsubscribe via the World Wide Web, visit
[Only registered users see links. ]
or, via email, send a message with subject or body 'help' to
[Only registered users see links. ]

You can reach the person managing the list at
[Only registered users see links. ]

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Yeast digest..."


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" <[Only registered users see links. ]>
Subject: [Yeast] Slow galactose induction
To: [Only registered users see links. ]
Message-ID:
<3fa3f1ca0709041054o692505aek8f75bbd8b3d1d072@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 [Only registered users see links. ].
Thanks,
Kerry



------------------------------

Message: 2
Date: Tue, 4 Sep 2007 15:27:43 -0600
From: "Z.J. Zhang" <[Only registered users see links. ]>
Subject: RE: [Yeast] Slow galactose induction
To: "Kerry Geiler" <[Only registered users see links. ]>, <[Only registered users see links. ].indiana.edu>
Message-ID:
<C9C1AF307F12AF4087DEF87CB0E6A4AD01A3881F@TELEGRAP H5.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
[Only registered users see links. ]
[Only registered users see links. ]



Reply With Quote
Reply

Tags
galactose , geiler , induction , kerry , slow


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
[Yeast] Slow galactose induction Z.J. Zhang Yeast Forum 0 09-04-2007 09:27 PM
Slow galactose induction Kerry Geiler Yeast Forum 0 09-04-2007 05:54 PM
Yeast stop to grow after galactose induction Nicolas Brandes Yeast Forum 0 03-04-2006 10:18 PM
It's looking bad for Bush & good for Kerry & the world! know buddee Physics Forum 0 10-28-2004 03:32 AM


All times are GMT. The time now is 08:46 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12408 seconds with 16 queries