Peter,
This issue seems to be quite a drawback.
I'd need a protease that is highly specific and constitutionally
active under cytosolic conditions but has virtually no targets in
unmodified cells. Any ideas what protease would be a good choice?
Wolfgang
>> > I imagine he wants to make it in bacteria to use for cleaving GST
> > tags off of fusion proteins. It very expensive to buy, aty least
> > it used to be..
>> I completely forgot about that use. If that is the case, I would
> say "find another enzyme". Although factor X can be easily
> activated by Russel's Viper Venom to form factor Xa, one must also
> purify (from rvv) and lyophilize the protein. Factor Xa is
> autoprotolytic and so this must be done fast. If your lab is not
> set up for large scale protein purifications, this would be
> difficult. As for Factor Xa, it has a very poorly defined
> recognition site. I can give you no less than 15 such sites that
> difer from the fXa thrombin cut site.
>> Peter Pediaditakis
Dr. Wolfgang Schechinger, Pathobiochemistry Dept.
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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Dr. Wolfgang Schechinger, Pathobiochemistry Dept.
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-----
*unsolicited mail is *NOT* appreciated, usual disclaimers apply
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