LOOKING FOR TIPS AND HINTS

Cornelius Krasel krasel at wpxx02.toxi.uni-wuerzburg.de
Tue Sep 1 15:32:49 EST 1998


Michael Vork <michael.vork at fys.unimaas.nl> wrote:
> I would like to have as many personal experiences as possible concerning the 
> production and purification of recombinant eukaryotic protein in E. Coli. 
> Which system is best and why (6xHis, GST, pET, IMPACT, other ??????)

It depends on your protein and what you want to do with it. For example, with
GST fusion proteins you can achieve superior purity compared to 6xHis, however,
you cannot purify proteins under denaturing conditions. Your protein may be
expressing in one of the systems but not in the other (or it may form
inclusion bodies). There may be steric hindrance in your assays if you put a
large domain into the wrong place. Etc. etc.

--Cornelius (working with 6xHis, GST, pET).

-- 
/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
/* "Science is the game we play with God to find out what His rules are."  */



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