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Fluorescence Resonance Energy Transfer as a Probe for G‐Quartet Formation by a Telomeric Repeat
Author(s) -
Mergny JeanLouis,
Maurizot JeanClaude
Publication year - 2001
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20010202)2:2<124::aid-cbic124>3.0.co;2-l
Subject(s) - förster resonance energy transfer , oligonucleotide , chemistry , fluorescence , folding (dsp implementation) , intramolecular force , circular dichroism , quenching (fluorescence) , fluorescence in the life sciences , covalent bond , guanine , resonance (particle physics) , photochemistry , dna , crystallography , stereochemistry , biochemistry , nucleotide , physics , organic chemistry , particle physics , quantum mechanics , electrical engineering , gene , engineering
The secondary structure of guanine‐rich oligodeoxynucleotides has been investigated with fluorescent probes. Intramolecular folding of a telomeric oligonucleotide into a quadruplex led to fluorescence resonance energy transfer (FRET) between a donor (fluorescein) and an acceptor (tetramethylrhodamine) covalently attached to the 5′ and 3′ ends of the DNA, respectively. Depending on oligonucleotide length, quenching efficiency varied between 0.45 and 0.72 at 20 °C. The conjugation of the dyes to the oligonucleotide had a limited, but significant, influence on the thermodynamics of G‐quartet formation. Intramolecular folding was demonstrated from the concentration independence of fluorescence resonance energy transfer over a wide concentration range. Folding of the oligonucleotide was confirmed by UV absorption, UV melting, and circular dichroism experiments. The folding of the G‐quartet could be followed at concentrations as low as 100 p M . Fluorescence resonance energy transfer can thus be used to reveal the formation of multistranded DNA structures.