Fusion proteins with CBP (calmodulin binding protein)

Giampaolo Minetti minetti at omni.cc.purdue.edu
Fri Sep 27 09:41:47 EST 1996


Hello,
I was wondering if anybody has ever tried the new family of expression vectors
Stratagene is now advertising under the name of Affinity System.
The vectors are deriver from the pET-11 series (T7 RNA polymerase, T7/LacO
promoter etc...). The recombinant protein is expressed as a fusion protein
to calmodulin binding protein (either C-terminal or N-teminal) and the resin
used for purification is a calmodulin affinity resin. They claim the
purification conditions be very mild (binding in the presence of Ca++,
elution with EGTA). I am thinking that handling such an affinity
resin might be somewhat troublesome (not easy to regenerate, proteolysis,
denaturation of calmodulin ...)
Any comments would be greatly appreciated
Thank you

Giampaolo Minetti
Purdue University
West Lafayette
IN
USA




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