microheterogeneity and tagged proteins

Cornelius Krasel krasel at alf.biochem.mpg.de
Sun Mar 20 15:25:29 EST 1994


Alan H. Schulman (schulman at convex.operoni.helsinki.fi) wrote:
: I would like to express a protein in E. coli for ultimate crystallization.
: It would be convenient to express it as a fusion protein.  Of the several
: systems available (6xHis (InVitroGen), strep-tag (Biometra),clip (Promega)
: etc., does anyone have experience with the result in crstallization?  How
: cleanly can the tag be cleaved off, without generating heterogeneity from
: multiple cleavage positions?

When I was a student, I once worked with antibody fragments expressed in
the periplasm of E. coli and tagged with a self-made his-tag. This tag
wasn't cleaved off at all and apparently did not interfere with
crystallization. Of course, you probably can't generalize from this
experience :-)

--Cornelius.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795 */
/* "People are DNA's way of making more DNA." (Edward O. Wilson, 1975)    */




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