pET expression help needed

mccallo at rand mccallo at rand
Fri Jul 22 11:07:04 EST 1994


Clemens Suter-Crazzolara <un691cs at genius.embnet.dkfz-heidelberg.de> writes:
> In your posting you write:
> >
> >hello, dear colleagues,
> >
> >is it possible to put the t7 tag used in the pET system at THE C-terminus
> >of a protein ? is it then still recognized by the corresponding antibody ?
> >i noticed that novagen offers the HIS TAG both at the N- and C- terminus,
> >but the T7 tag only at the N-terminus.
> >
> >thanks, clemens
> >
> >
>
> T7 isn't a tag. It's a promoter element recognized by T7 RNA polymerase
> which is required....

Sorry Clemens, but you're mistaken; check your Novagen catalog.  Novagen sells
a monoclonal Ab directed against the 11 amino acid gene 10 leader peptide. 
They call this peptide the " T7-Tag", which is what the poster was refering to.

To the original poster:  Yes, it will work at either end, as long as that end
is not buried internally within the folded protein;  it must be available at
the surface of the protein.  Depends upon the higher order structure of the
protein;  denaturing the protein will probably make it available in any case,
if that is acceptable for your purposes.  

Owqen McCall
Abbott Laboratories
Abbott Park, IL USA
mccall.owen at igate.pprd.abbott.com
  



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