Taq polymerase patent (not)

[Tom Bickle]

In article <graham.839119732 at biodec.wustl.edu>, graham at biodec.wustl.edu
(Dr. James E. Graham) wrote:

> > I always thought patent only apply if you want make money out of it.  I
> > thought that anyone can make the enzyme "at home" even for PCR, as long
> > as he doesn't sell it. Isn't that true?
> 
> A recent Biotechniques article describes a primer pair, and a method 
> where one can clone the Taq pol gene from T. aquaticus DNA by PCR,
> express it in E.coli, and purify it in one step by simply boiling the 
> cell lysate, as the polymerase is then apprenlty the only enzyme in the 
> cell which does not denature, aggregate and precipitate. Haven't 
> tried it myself.
> 
> A literature search should point the way. :)
> 
> Jim
> J. Graham PhD 

However, a boiled E. coli extract also contains other proteins, including
HU, which is a DNA binding protein. I would be cautious about using the
boiled prep as is. A simple gel filtration step would be helpful as all
the thermostable E. coli proteins are small.
> Biology Department

-- 
Tom Bickle, Department of Microbiology, Biozentrum, Basel University
Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
+4161 267 2120, Fax: +4161 267 2118  bickle at ubaclu.unibas.ch