(His)6 tags for protein purification

Tony Houthaeve houthaeve at embl-heidelberg.de
Tue May 10 11:43:34 EST 1994


In article <DRESNICK.93Oct13102645 at m14s-010-4.mit.edu>, dresnick at athena.mit.edu (David I Resnick) writes:
> Hi,
> 
> Has anyone out there used the Hisx6 tags for purification of recomb.
> expressed proteins?  I'm considering putting it on a fragment of a
> protein we are studying and expressing it in yeast.  However I have
> the following questions/concerns:
> 
> 1) Does the His6 tag need to be at the very N or C terminus for the 
>    protein for the protein to bind to the Ni2+ column?  Or can
>    it be internal as long as it is solvent exposed?
> > --
>        David Resnick                           dresnick at athena.mit.edu      

We've made here overexpressed proteins (=vaccinia expressed Hela cells)
of for instance SRF and many more proteins (mainly in the group of 
Henk Stunnenberg here at the EMBL).
There does not seem to be any problem tagging the His-tag to the N, 
neither to the C-terminus. As long as the tag does not interfere with
functionality. I would be sceptic on adding the tag in the protein, as the
structure might change drastically.
Any way, before starting the project try to predict the 3-D structure of the
protein and see if the modification is allowed [sic] or not.

Good luck








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