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001-es BibID:BIBFORM004641
Első szerző:Vámosi György (biofizikus)
Cím:The helix-coil transition of DNA duplexes and hairpins observed by multiple fluorescence parameters / Vamosi, G., Clegg, R. M.
Dátum:1998
ISSN:006-2960 (Print)
Megjegyzések:The thermal denaturation of 8-20-bp DNA duplexes labeled with fluorescein and tetramethylrhodamine at opposing 5'-ends was investigated by monitoring the fluorescence intensity of the dyes, the fluorescence anisotropy of tetramethylrhodamine, the fluorescence resonance energy transfer between fluorescein and rhodamine, and, for the 20-bp duplex, the UV absorption. Melting experiments with the single strands of the duplexes revealed that the single strands can form hairpins stabilized by only a few base pairs. The thermal denaturation curves of the duplexes were fitted well to an extended all-or-none model assuming that only the fully base-paired duplex, the maximally base-paired hairpin, and the random coil conformations are present simultaneously. The extent-of-melting versus temperature curves derived from the different spectroscopic parameters are nearly identical, provided that the analysis of the baselines is carried out correctly; the DeltaH and DeltaS of the dissociation compare well with predictions based on nearest neighbor interaction values available in the literature. Our results imply that for all the oligonucleotides other than the 34-bp oligomer, no partially melted intermediates other than hairpins are present in the reaction mixture in amounts that can be detected by our methods. The melting of the hairpins was also studied directly using single-stranded oligonucleotides. The melting of a 34-bp duplex can be accounted for by a statistical zipper model
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
analysis
chemical synthesis
chemistry
Dna
Single-Stranded
Dyes
Energy Transfer
Entropy
Fluorescein
Fluorescence
Fluorescence Polarization
Fluorescence Resonance Energy Transfer
Fluorescent Dyes
Heat
methods
Models,Chemical
Nucleic Acid Conformation
Nucleic Acid Denaturation
Nucleic Acid Heteroduplexes
Oligonucleotides
Research
Rhodamines
Spectrometry
Support
Temperature
Thermodynamics
Megjelenés:Biochemistry. - 37 : 40 (1998), p. 14300-14316. -
További szerzők:Clegg, Robert M.
Internet cím:elektronikus változat
DOI
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