GST fusions - please, help

Armin Sepp Armin.Sepp at bbsrc.ac.uk
Tue Oct 24 08:18:52 EST 1995


krasel at wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) wrote:
>ijiwaru at wheel.dcn.davis.ca.us wrote:
>> In article <46bbgc$16vi at news.doit.wisc.edu>, klenchin at macc.wisc.edu (Dima
>> Klenchin) wrote:
>
>> > I need to overexpress 70K protein as a GST fusion. It is cloned into
>> > Pharmacia's pGEX-2T. As usually, expression level and solubility is a
>> > problem. 
>> > 
=========cut
>A funny thing with BL21 seems to be that it might be necessary to
>screen clones for expression levels. I am expressing another protein
>in BL21 (actually BL21(DE3)pLysS, but I don't think that matters);
>I tested four clones which expressed the protein in widely different
>ranges.
============cut============
>--Cornelius.
>
Dear Cornelius

Did you take any measures to keep the pLysS plasmid in the bug? It carries a 
chloramphenicol resistance gene.

with best wishes

-- 
* Armin Sepp, PhD           \                                    *
* Department of Immunology   \     "Every little helps." (Tesco) *
* Babraham Institute          \                                  *
* Cambridge CB2 4AT,U.K.       \      Armin.Sepp at bbsrc.ac.uk     *





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