enterokinase

Per Mygind perm at biobase.dk
Fri Oct 22 06:17:49 EST 1999


Alexandre wrote:

> hi everybody,
>
> I want to express a protein linked to a very soluble fusion partner. To
> remove the fusion partner at the end of the purification, I want to
> engineer a bovine enterokinase site, which cleaves (specifically ?!?) a
> given sequence.
> Does anybody have experience with that protease ? which one (from which
> supplier) performs best ? do you have any idea about the conditions
> (ions, pH...) I have to use to get an efficient cleavage.
>
> thanx for any help
>
> Alexandre Mooser
> Department of Biochemistry
> University of Zurich
> Switzerland

Enterokinase is a poor protease, we often get unspecific cleavage, the
protein is not all cleaved and it is expensive.
 In our hands, Thrombin is a much more reliable, cheaper and more
effective protease.
We have successfully incoorporated the recognitionsite (6aa) in many of
our fusion protein
constructs. We use Thrombin from pharmacia, no affiliation by the way.

--
Regards

Per Mygind

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If you are are not part of the solution, you are part of the precipitate
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Per Mygind, Cand.scient

Department of Medical Microbiology and Immunology
The Bartholin Building, University of Aarhus, Denmark
phone : 89 42 17 47, fax   : 86 19 61 28

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