Comparing mono- and divalent DNA groove binding cyanine dyes--Binding geometries, dissociation rates, and fluorescence properties
Journal article, 2006

The unsymmetrical cyanine dyes BOXTO-PRO and BOXTO-MEE were derived from the DNA groove binder BOXTO, by adding a positively charged or a non-ionic hydrophilic tail to BOXTO, respectively. The main objective was to obtain more efficient DNA probes, for instance in electrophoresis and microscopy, by slowing down the dissociation of BOXTO from DNA. The interactions with mixed sequence DNA was studied with fluorescence and absorbance spectroscopy, stopped-flow dissociation and gel electrophoresis. Both the derivatives are groove bound as BOXTO, and have similar fluorescence properties when bound to mixed sequence DNA in free solution. BOXTO-PRO exhibits a slower dissociation than BOXTO from DNA, whereas the dissociation rate for BOXTO-MEE is faster and, unexpectedly independent of the ionic strength. During gel electrophoresis both BOXTO-PRO and BOXTO-MEE exhibit a faster dissociation rate than BOXTO. Still, BOXTO-PRO seems to be a good alternative as DNA probe, especially for applications in free solution where the dissociation is slower than for the corresponding intercalator TOPRO-1.

Cyanine dyes

Dissociation kinetics

Groove binding to mixed sequence DNA

Spectroscopy

Gel electrophoresis

Author

Maja Eriksson

Chalmers, Chemical and Biological Engineering

Fredrik Westerlund

Chalmers, Chemical and Biological Engineering

Merima Mehmedovic

Chalmers, Chemical and Biological Engineering

Per Lincoln

Chalmers, Chemical and Biological Engineering

Gunnar Westman

Chalmers, Chemical and Biological Engineering

Anette Larsson

Chalmers, Chemical and Biological Engineering, Pharmaceutical Technology

Björn Åkerman

Chalmers, Chemical and Biological Engineering

Biophysical Chemistry

0301-4622 (ISSN)

Vol. 122 3 195-205

Subject Categories

Physical Chemistry

DOI

10.1016/j.bpc.2006.03.009

More information

Latest update

11/5/2018