Photophysical characterization of a FRET system using tailor-made DNA oligonucleotide sequences.
We have carried out a detailed photophysical study of the FRET D/A pair consisting of a carbostyril donor and a Ru(II)bathophenanthroline complex acceptor in double-stranded synthetic DNA. Altogether 13 different double-stranded 30 base pair DNAs showing small incremental differences in the distances between donor and acceptor were synthesized. Using the fluorescence of the donor as well as of the acceptor, D/A separations were determined and compared to those derived from a well-established model for DNA distance calculations. The model calculations and anisotropy studies revealed that the donor can nearly be seen as a free rotator allowing the application of the established FRET data evaluation.
University of Potsdam, Institute of Chemistry (Physical Chemistry), 14476 Potsdam-Golm, Germany.
Roman
R
Flehr
Andrea
A
Kienzler
Willi
W
Bannwarth
Michael U
MU
Kumke
D001483:Base Sequence; D004247:DNA / Q000138:chemical synthesis / Q000378:metabolism*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D006912:Hydroxyquinolines / Q000378:metabolism*; D008958:Models, Molecular; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D009942:Organometallic Compounds / Q000378:metabolism*; D010618:Phenanthrolines / Q000378:metabolism*; D056951:Photoelectron Spectroscopy; D015363:Quinolones / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D006912:Hydroxyquinolines; D009841:Oligonucleotides; D009942:Organometallic Compounds; D010618:Phenanthrolines; D015363:Quinolones; C010064:carbostyril; D004247:DNA; C041132:tris(4,7-diphenyl-1,10-phenanthroline)ruthenium (II)