Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Aryeh,

yes, in my hands mYFP-mRFP1 was giving weaker (ca. 10-fold as to mCeFP-mYFP) 
but readily detectable signal in sensitized FRET measurements according to 
Gordon et al. (1998). In contarst to mCherry, mRFP1 is not tagged with 
GFP-derived peptides.

Lily, if you have time and enjoy drinking Champagne afterwards, go ahead and 
make the mCherry more "viable", shorter at the N-terminus. I will give you 
the hinge sequnce on the basis of limited confidentiality.

Cheers,

Vitaly


----- Original Message ----- 
From: "Aryeh Weiss" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Tuesday, July 17, 2007 1:20 PM
Subject: Re: eYFP-mCherry Fret


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Ammasi Periasamy wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Hello
>>
>> We tried the mOrange with eYFP, it works ok, not great. Most of the red 
>> proteins are not working good for FRET, do not know why. Some one tried 
>> mCherry, it did not work. If I get more info, I will put it in the list 
>> server.
>>
> I have measured 30-40% FRET efficiency with a YFP-mRFP1 fusion, using the 
> acceptor photobleaching method. Given the minimum distance between the 
> fluorophores which is imposed by the protein structure, and the likelyhood 
> that the alignment of the dipoles is not perfect,  I would not expect 
> more.
>
> --aryeh
> -- 
> Aryeh Weiss
> School of Engineering
> Bar Ilan University
> Ramat Gan 52900 Israel
>
> Ph:  972-3-5317638
> FAX: 972-3-7384050
>