Seeking for a FRET pair

[yee z]

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

Hi Everyone,

 

I start a new job in the imaging core recently, and think to give a demo show on how fret works in our new live cell imaging system.

 

I would highly appreciate if anyone on this list can provide me an easy working FRET pair like plasmids that I can transfect cell with or let me know where I can purchase such things.

 

 

PS: I am located in Boston, USA.

 

Thanks,

 

Yi

 

 

[George McNamara]

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

mTFP (mTFP1) to mVenus or mKO looks good. mTFP is a better match for a 457nm laser line than Cerulean and does not have some of Cerulean's funky behavior (described in another article in the JBO issue below). For mTFP -> Venus see:


http://spiedl.aip.org/getabs/servlet/GetabsServlet?prog=normal&id=JBOPFO000013000003031203000001&idtype=cvips&gifs=Yes

J. Biomed. Opt. / Volume 13 / Issue 3 / <a href="http://spiedl.aip.org/dbt/dbt.jsp?KEY=JBOPFO&amp;Volume=13&amp;Issue=3#Special Section on Visible Fluorescent Proteins"> Special Section on Visible Fluorescent Proteins

Characterization of an improved donor fluorescent protein for Förster resonance energy transfer microscopy

J. Biomed. Opt., Vol. 13, 031203 (2008); DOI:10.1117/1.2939094

Richard N. Day and Cynthia F. Booker
University of Virginia Health System, Departments of Medicine and Cell Biology, P. O. Box 800578, Charlottesville, Virginia 22908-0578

Ammasi Periasamy
University of Virginia, W.M. Keck Center for Cellular Imaging, Departments of Biology and Biomedical Engineering, Gilmer Hall, Charlottesville, Virginia 22904-4328
The genetically encoded fluorescent proteins (FP), used in combination with Förster resonance energy transfer (FRET) microscopy, provide the tools necessary for the direct visualization of protein interactions inside living cells. Typically, the Cerulean and Venus variants of the cyan and yellow FPs are used for FRET studies, but there are limitations to their use. Here, Cerulean and the newly developed monomeric Teal FP (mTFP) are compared as FRET donors for Venus using spectral and fluorescence lifetime measurements from living cells. The results demonstrate that when compared to Cerulean, mTFP has increased brightness, optimal excitation using the standard 458-nm laser line, increased photostability, and improved spectral overlap with Venus. In addition, the two-photon excitation and fluorescence lifetime characteristics are determined for mTFP. Together, these measurements indicate that mTFP is an excellent donor fluorophore for FRET studies, and that its use may improve the detection of interactions involving proteins that are difficult to express, or that need to be produced at low levels in cells.

Ai HW, Olenych SG, Wong P, Davidson MW, Campbell RE.
Hue-shifted monomeric variants of Clavularia cyan fluorescent protein: identification of the molecular determinants of color and applications in fluorescence imaging.
BMC Biol. 2008 Mar 6;6:13.
PMID: 18325109 [PubMed - indexed for MEDLINE]

Ai HW, Henderson JN, Remington SJ, Campbell RE.
Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging.
Biochem J. 2006 Dec 15;400(3):531-40.
PMID: 16859491





At 09:53 AM 7/10/2008, yee z wrote:

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

Hi Everyone,

 

I start a new job in the imaging core recently, and think to give a demo show on how fret works in our new live cell imaging system.

 

I would highly appreciate if anyone on this list can provide me an easy working FRET pair like plasmids that I can transfect cell with or let me know where I can purchase such things.

 

 

PS: I am located in Boston, USA.

 

Thanks,

 

Yi

 

 

 

George McNamara, Ph.D.
University of Miami, Miller School of Medicine
Image Core
Miami, FL 33010
[hidden email]
[hidden email]
305-243-8436 office
http://home.earthlink.net/~pubspectra/
http://home.earthlink.net/~geomcnamara/
http://www.sylvester.org/research/SR_lab_analytical.asp?ana=desc (Analytical Imaging Core Facility)