pET expression

gcagney gcagney at indigo.ie
Thu Mar 13 17:38:57 EST 1997


We've been using the Novagen pET system to express buckets of His-tagged 
recombinant proteins in E.coli.  The proteins form inclusion bodies and are 
recovered using affinity chromatography under denaturing conditions 
(guanidine or urea).  The problem is removing the denaturant...there is so 
much protein that it inevitably precipitates during dialysis.  

What is the secret of avoiding precipitation?

Thanks,

Gerard Cagney PhD
Elan Pharmaceutical Technologies, Biotechnology Building, Trinity College, 
Dublin 2, Ireland
Tel: +353 1 6710901
Fax: +353 1 6710920
Email: gerardc at iol.ie




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