E coli expression of eucaryotic protein

Brad Nicholson brad at corona.med.utah.edu
Tue Jul 18 19:45:03 EST 1995


In article <kallisti-1707952017580001 at lomasneylab.feinberg.nwu.edu>, 
kallisti at merle.acns.nwu.edu (Patrick Grealish) wrote:

> HEllo, I am current starting another experiment where i'll need to 
produce
 >eucarotic protein in E. coli. 
>snip<
Hello Patrick,
Our lab uses the GST system from Pharmacia.  You get a fusion protein with 
Glutathione-S-Transferase and can cleave with your choice of proteases.  I 
have used pGEX-2T with success, although I have had problems getting my 
protein cleaved away from the GST.  I didn't need my protein absolutely 
pure, so it wasn't important.  Another person in the lab has used pGEX-2T &
pGEX-2TK with good success.  The latter vector is usefull in that it has a 
protein kinase site after the protease site, but in front of your protein 
that you can lable stinking hot with gamma 32 ATP.

Good Luck
Brad
===========================================================================
Brad Nicholson              |"If it worked the first time, it wouldn't be 
Department of Pathology     |	research."...Brad Nicholson
University of Utah          |	Live from behind the Zion Curtian.
Salt Lake City, UT 84132    | 
brad at corona.med.utah.edu    |
or: (801)-581-4365          | My opinions are solely my own.
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