Johan Svennilson wrote:
>> I would like to know the effects of NH- or COOH- location of GFP on a
> target protein to the property of the target protein, especially the
> target protein is a membrane receptor. If anybody has some experience
> with this?
>> Thank you for your help!
>> Xinyu Wang
> post-doc at Karolinska Institut,
> please reply to: wanxi at child.ks.se
N-terminal tagging may interfere with trafficking especially if large
N-termini are located to the extracellular space. If signaling peptides
are present, one may try to insert the GFP at the end of the signal
peptide. But be aware that the exact boundaries are sometimes a matter
of speculation and may be even changed due to insertion of GFP.
C-terminal tagging is less critical but nedds the removal of stop codon
of you protein of interest. Most promising: try both and check the
funcional integrity of your receptor.
Michael Schaefer
Inst. fuer Pharmakologie
Freie Universitaet Berlin
