elution of GST fusions

NOTHWEHR at BIOSCI.MBP.MISSOURI.EDU NOTHWEHR at BIOSCI.MBP.MISSOURI.EDU
Sat Jun 21 22:36:17 EST 1997


We have successfully expressed a GST fusion protein in E. coli.  The
protein of interest is not fused to the C-terminus of GST as is usually
the case but rather to the N-terminus. Our problem is that we have not
been able to elute the fusion protein from glutathione agarose using
reduced glutathione even after prolonged incubation periods at room
temp.  In contrast GST alone elutes just fine under the conditions we
have used.  Of course our fusion protein does elute under denaturing
conditions such as 100 deg. in the presence SDS-PAGE laoding buffer. 
Does anyone know of any other conditions involving reduced glutathione
or a related molecule that would specifically dissociate GST fusions
from glutathione agarose?  Thanks.



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