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Journal of Biomedical Optics

Photophysical properties of Cerulean and Venus fluorescent proteins
Author(s): Pabak Sarkar; Srinagesh V. Koushik; Steven S. Vogel; Ignacy Gryczynski; Zygmunt Karol Gryczynski
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Paper Abstract

Cerulean and Venus are recently developed fluorescent proteins, often used as a donor-acceptor pair by researchers in Förster resonance energy transfer-based colocalization studies. We characterized the fluorescent properties of these two proteins in a broad spectral range (form ultraviolet to visible region). Excitation spectra, lifetimes, and polarization spectra show significant energy transfer from aromatic amino acids to the fluorescent protein chromophore. High steady-state anisotropy values and the lack of a fast component in anisotropy decays show that the fluorescent protein chromophore is rigidly fixed within the protein structure. Furthermore, we show that the chromophores are not accessible to external quenchers, such as acrylamide or potassium iodide (KI), allowing the removal of "unwanted" background in the environment with external quencher, while leaving the Cerulean/Venus fluorescence unchanged.

Paper Details

Date Published: 1 May 2009
PDF: 9 pages
J. Biomed. Opt. 14(3) 034047 doi: 10.1117/1.3156842
Published in: Journal of Biomedical Optics Volume 14, Issue 3
Show Author Affiliations
Pabak Sarkar, Univ. of North Texas Health Science Ctr. at Fort Worth (United States)
Srinagesh V. Koushik, National Institutes of Health (United States)
Steven S. Vogel, National Institutes of Health (United States)
Ignacy Gryczynski, Univ. of North Texas Health Science Ctr. at Fort Worth (United States)
Zygmunt Karol Gryczynski, Univ. of North Texas Health Science Ctr. at Fort Worth (United States)


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