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Hi Lily,

this pair does not work well, as R.Tsien's people added the GFP-derived 
peptides to both N- and C-terminus of the mCherry.

It looked like they were struggling with something like solubility of the 
protein.

However, if you remove the N-terminal peptide including the Met10 within the 
fusions with the the C-terminus of the protein of ineterest, then the 
monomeric YFP (F64L, A206K, L221K, F223R) would be a great donor and the 
"truncated" mCherry a reasonable acceptor (it is very unlikely that is 
monomeric in vivo, though I see less aggregation with a number of protein 
fusions in comparison to mRFP1 - a true dimer/oligomer in vivo!!! or to mYFP 
mentioned above). Make sure you have a relatively long Gly-rich flexible 
"natural" hinge, let say of SV40 origin - it is known among the older 
generation of crystalographers at Scripps.

Good luck,

Vitaly

301-846-6575


---- Original Message ----- 
From: "Koo, Lily (NIH/NIAID) [F]" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Tuesday, July 17, 2007 7:37 AM
Subject: eYFP-mCherry Fret


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Greetings All,
>
> One of our users would like to try the FRET pair eYFP-mCherry.  Does
> anyone know if this pair will work?
>
> Thank you,
>
> Lily
>