Yeast Digest, Vol 45, Issue 2

Hello, Everyone,

FCY2 gene is an important purine-cytosine permease. However, there two more putative purine-cytosine permeases encoded by FCY21 and FCY22. But there is lack of information on exact substrates and regulation of their activity etc. Only information I get is: Fcy21p and Fcy22p are very similar to Fcy2p but can not substitute its function.

I wonder if anyone have more information on FCY21 and FCY22 and their protein product functions. Any pointers are appreciated.

Thank you

Shaun

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Today's Topics:

1. RE: Yeast Digest, Vol 45, Issue 1 (Andrew Carter)


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Message: 1
Date: Wed, 4 Feb 2009 09:27:26 -0800
From: "Andrew Carter" <[Only registered users see links. ].edu>
Subject: [Yeast] RE: Yeast Digest, Vol 45, Issue 1
To: <[Only registered users see links. ].indiana.edu>, <[Only registered users see links. ].indiana.edu>
Message-ID: <000001c986ed$daa298f0$8fe7cad0$@ucsf.edu>
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Hi Thomas
Glucose represses the Gal promoter whereas Raffinose doesn't. If you try
growing up yeast in YPD (2% glucose) and then add galactose you will get no
(in my experience) induction. If you grow up in YP raffinose and add
galactose the promoter will get turned on immediately.

It turns out however that you can first grow in glucose and then add
galactose. You just have to make sure that you have used up all your
glucose. I have been playing with this quite a lot recently (in a large
fermentor) and it works well. The key for me was using 2X YP and 1% glucose
and waiting until growth plateaus. Then add in 2% galactose (0.5% and 1%
also work) and induction turned on just fine. I was using induction from a
gal promoter inserted in the genome and haven't tried it with a plasmid
based system, but my guess is it should work. The real advantage is that
Raffinose gets really expensive if you want to scale up...

Not sure why growth is slower in galactose than glucose but I see it alot
(final OD is usually lower as well).
Best wishes
Andrew






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Today's Topics:

1. raffinose metabolism (Thomas, Arun)


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Message: 1
Date: Wed, 4 Feb 2009 11:36:10 +0100
From: "Thomas, Arun" <[Only registered users see links. ]-Osnabrueck.DE>
Subject: [Yeast] raffinose metabolism
To: <[Only registered users see links. ].indiana.edu>
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Hi everyone,

To induce a gene under the GAL promoter from a 2micron plasmid, why is it
preferred to grow the cells in raffinose/glucose initially followed by
induction with galactose?

If cells grow slowly in galactose, why is it so?

Thank you,
Thomas



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