GST Fusion Proteins in Yeast
dan at cubsps.bio.columbia.edu
Sat Jul 8 10:02:26 EST 1995
In article <3tkgd9$pt at news.service.uci.edu>,
Bill Brickey <wwbricke at uci.edu> wrote:
> 1. Is there something in yeast extracts which interferes with
> binding? Are there alternate protocols ?
I don't think there is any overall problem in yeast. People in my lab have
expressed GST fusions in yeast, but purification was limited by the lower
There may be other explanations. A protein I have a GST fusion with seems
to stick very tightly to my column (Pharmacia pre-packed). Our procedure
called for elution with 1mM glutathione but I found that it wasn't coming
off. I thought it was not sticking, but when I used more glutatione (I now
elute with 20mM in 7 column volumes) a ton came tumbling off the column.
In earlier experiments I didn't see the protein in the high salt washes we
do at the end either, which is strange.
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