His tag.

Michael Witty mw132 at mole.bio.cam.ac.uk
Sat Mar 17 07:15:06 EST 2001


On Fri, 16 Mar 2001, Frank Fuerst wrote:

> klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu (Dima Klenchin) wrote:
> 
> >skumar at mbu.iisc.ernet.in ("S. Kumar Singh") wrote:
> >:Hi all,
> >:Is there any good established procedure for removing His tag from the N
> >:terminus of a protein., I would greatly appreciate a reference on this.
> >:Thanks a million in advance.
> >
> >Only if you have protease recognition site between the tag and
> >the protein. Most conventional proteases are not specific enough
> >and cut expressed protein as well, however. TEV protease and
> >its 6 aa recognition site appears to be the best choice. Look up
> >Gibco catalog for more details.
> 
> Deep down in my mind there is something about inteins (self-splicing
> proteins) with a his-tag, fused to the protein of interest: Bind it to
> a Ni-column, induce splicing activity and elute clean, tag-free
> protein; then clean your column by eluting the remaining intein with
> imidazole. Unfortunately, I don't remember a manufacturer name or
> reference.
> 
> Bye, Frank

Frank, this sounds like New England Biolab's intein technology.  There is
a lot about it on their web site and catalogue.  Mike.




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