mTFP might be a good donor, however based on my experience, v
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Hi Mike,

mTFP might be a good donor, however based on my experience, venusYFP is not the 
best acceptor for the FRET studies. EYFP with the F64L substitution worked 
better for me. ...though in the very beginning I was in a "blind" love with the 
venusYFP too. Also, the paper did not show any advantage of mTFP over the CeFP. 
In addition, though the CeFP carries A206K substitution, I would recommend to 
add well known monomeric mutations in positions 221 and 223 to CeFP and all 
three to EYFP(F64L). 
As it is difficult to monitor fluorophore concentration on a voxel-by-voxel 
basis, highly accurate, quantitative, comparative FRET measurements study is 
still a challenging task (FLIM inclusive). George, do you have a good reference 
to?

As to the vectors, just do it yourself - it is very simple and quick, PCR is a 
powerful tool and Dave Piston is the best source for the CeFP(A206K). 

Good luck,

Vitaly
301-515-7833




________________________________
From: George McNamara <[log in to unmask]>
To: [log in to unmask]
Sent: Fri, January 28, 2011 9:32:26 PM
Subject: Re: CFP YFP plasmid vectors

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Contact Richard Day, Indiana Univ, ask for the constructs published in:

Characterization of an improved donor fluorescent protein for Forster 
resonance energy transfer microscopy. </pubmed/18601527>

Day RN, Booker CF, Periasamy A.

J Biomed Opt. 2008 May-Jun;13(3):031203.PMID: 18601527

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2483694/?tool=pubmed



On 1/28/2011 11:18 AM, Mike Tighe wrote:
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> To join, leave or search the confocal microscopy listserv, go to:
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>
> Would anyone be willing to recommend a source for YFP and CFP plasmid
> vectors for FRET, preferably compatible with D5Hα. Any feedback would be
> most appreciated.
>
> Thanks!!
> Mike
>
>
>
>
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-- 


George McNamara, PhD
Analytical Imaging Core Facility
University of Miami