GROEL and GST

didier d-fesque at nimr.mrc.ac.uk
Tue Jul 8 13:43:58 EST 1997


about your question!


I am coexpressing GroEL and GroES with a GST fusion protein. 
The GroEL seems to copurify with my desired GST fusion protein on the
glutathione sepharose 4B column. Has anyone experienced this?
Any suggestions?

Kelley Ruud
University of Texas at Austin

Hi,

I think you will have problem to get rid of the chaperone protein while
keeping your GST protein soluble. nevertheless, I have seen in the GST
booklet from pharmacia that you can get rid of DNAK chaperone which is
very often associated with GST with a cocktail of ATP/MgCl. I remenber a
PNAS paper a few years ago (one of the author is stieger) describing the
coexpression of a kinase with GROEL, using a special buffer they succeed
to get their protein soluble without GROEL/GROES! have a look in
medline.
IF you are successful pleased tell me
  
good luck


didier



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