Malcolm M. Campbellcampbell
campbell at cict.fr
Fri Feb 5 07:11:53 EST 1993
I realize that the following question is becoming a FAQ (and I'm kicking
myself for not saving previous postings) but...
What is the consensus out there regarding the best recombinant protein
expression system based on the following criteria:
1) system must produce a functional protein (ie. with catalytic activity);
2) protein is easily purified to _homogeneity_ AND in high quantities
(think mg's), and;
3) the cells carrying the recombinant protein are easily transformable
and can be grown using standard bacterial culture facilities?
We have already expressed active protein (from the cDNA encoding our plant-
derived protein) using the pT77 system - it works great if we want to assay
catalytic activity in crude preps. However, now we want pure protein for
NMR analysis of protein/substrate/inhibitor interactions and, of course,
crude preps just aren't going to cut it. After having developed a number
of protein purification protocols for this enzyme from a number of plant
species, we aren't too enthuastic about having to tackle the same problems
with our bugs. A nice, simple protocol would be great...
Can anyone out there help us?
Thanks in advance.
Malcolm M. Campbell + phone: (33) 61 55 69 05
Equipe Lignines + FAX: (33) 61 55 62 10
Centre de Biologie et Physiologie Vegetales + internet:
URA CNRS No. 1457 + campbell at cix.cict.fr
Universite Paul Sabatier +
118 route de Narbonne +
31062 Toulouse cedex +
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