Yeast as an alternative expression system

Kevin Pumiglia pumiglk at aa.wl.com
Fri May 5 08:08:07 EST 1995


In article <D82q5E.6Dw at phibred.com>, bipin dalmia <dalmiabk at phibred.com>
wrote:

> In article <pumiglk-040595104941 at dom006.dom.aa.wl.com> Kevin Pumiglia,
> pumiglk at aa.wl.com writes:
> >vector we are using were generously provided by Bert O'Malley's Lab at
> >Baylor (see Biotechniques, last Feb. maybe?).  One draw back of this
> 
> can you please post the reference? do you know if anyone is working on
> making a yeast vector with GST and thrombin cleavage?
> 
> bip

Biotechniques 16(2):194-196, 1994 is the description of the article I was
refering to.  I have constructed what I believe is a modification of this
vector with the insertion of the glycine kinker and thrombin cleavage site
from pGex-KG.  This vector however is still untested as it was only
recently done and I have not cloned in an insert protein and checked
cleavage.  When this is ready for prime time I would be happy to share. 
There also exists a vector constructed by Christopher Hugs at Washington
University (St. Louis, MO) which incorporates a factor Xa endopeptidase
site dowstream of GST.  This is driven by the Gal10 promoter and is has
several selection flavors e.g., Trp, His, Ura, and Leu.  These should be
available from ATCC.

--Kevin



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