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European Biophysics Journal : Volume 27, Issue 1, January 1998

European Biophysics Journal : Volume 26

European Biophysics Journal : Volume 25

Triple helix DNA oligomer melting measured by fluorescence polarization anisotropy

Content Provider	Springer Nature Link
Author	Barone, Flavia Chirico, Giuseppe Matzeu, Mirella Mazzei, F. Pedone, Francesco
Copyright Year	1998
Abstract	A synthetic DNA triple helix sequence was formed by annealing a pyrimidinic 21 mer single strand sequence onto the complementary purinic sequence centred on a 27 mer duplex DNA. Melting of the third strand was monitored by UV spectrophotometry in the temperature range 10–90 °C. The Tm of the triplex, 37 °C, was well separated from the onset of duplex melting. When the same triple helix was formed on the duplex bearing one nick in the center of the pyrimidinic sequence the Tm of the triplex was shifted to approximately 32 °C and overlapped the melting of the duplex. We have used fluorescence polarization anisotropy (FPA) measurements of ethidium bromide (EB) intercalated in duplex and triplex samples to determine the hydrodynamic parameters in the temperature range 10–40 °C. The fluorescence lifetime of EB in the samples of double and triple stranded DNA is the same (21.3 ± 0.5 ns) at 20 °C, indicating that the geometries of the intercalation sites are similar. The values for the hydration radii of the duplex, normal triplex, and nicked triplex samples were 10.7 ± 0.2, 12.2 ± 0.2, and 12.0 ± 0.2 Å. FPA measurements on normal triplex DNA as a function of temperature gave a melting profile very similar to that derived by UV absorption spectroscopy. For the triplex carrying a nick, the melting curve obtained using FPA showed a clear shift compared with that obtained for the normal triplex sample. The torsional rigidity of the triplex forms was found to be higher than that of the duplex form.
Starting Page	137
Ending Page	146
Page Count	10
File Format	PDF
ISSN	01757571
Journal	European Biophysics Journal
Volume Number	27
Issue Number	2
e-ISSN	14321017
Language	English
Publisher	Springer-Verlag
Publisher Date	1998-02-27
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Medicine Biophysics

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