Hi Katherine, Maybe you can try some useful ideas for your case listed in this study: Protein Expr Purif. 2007 April; 52(2): 280–285. Effect of Osmotic Stress and Heat Shock in Recombinant Protein Overexpression and Crystallization. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1865119/ Best wishes
Toufic El Arnaout Membrane Structural and Functional Biology Group Trinity College Dublin On Mon, Sep 24, 2012 at 3:38 PM, Katherine Sippel < [email protected]> wrote: > Hi All, > > I've recently swapped over to expression in the ArcticExpress(DE3) cells > for a particularly rock-like protein. I've got soluble expression but I'm > having the issue of Cpn60 (the overexpressed chaperonin) piggybacking along > with the tagged protein. Google-fu and the CCP4bb archives indicate that > this is a known issue but I haven't seen a solution thus far. Does anyone > out there have any tricks up their sleeve? > > Also in case you were wondering in the various BL21 lines, even at > extremely low temperatures and 10 uM IPTG, it expresses well but produces > inclusion bodies that are completely insoluble in 8M urea, 6M guanidine, > high temperature, high pH, high reducing agent, and a number of detergents > so swapping back to another cell line isn't in the cards. > > Any help would be appreciated. > > Cheers, > Katherine >
