期刊
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
卷 465, 期 1, 页码 90-100出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.abb.2007.04.034
关键词
fluorescent dyes; aggregation; T-jump; stopped-flow; calf-thymus DNA; intercalation
Kinetics and equilibria of cyanine dyes thiazole orange (TO) and benzothiazole orange (BO) self-aggregation and binding to CTDNA are investigated in aqueous solution at 25 C and pH 7. Absorbance spectra and T-jump experiments reveal that BO forms J-aggregates while TO forms more stable H-aggregates. Fluorescence and absorbance titrations show that TO binds to DNA more tightly than BO. TO stacks externally to DNA for low polymer-to-dye concentration ratios (C-P/C-D) while dye intercalation occurs for high values of CP/CD. T-jump and stopped-flow experiments performed at high CP/CD agree with reaction scheme D+S reversible arrow D,S reversible arrow: DSI reversible arrow DSII where the precursor complex D,S evolves to a partially intercalated complex DS, which converts to the more stable intercalate DSII. Non-electrostatic forces play a major role in D,S stabilization. Last step is similar for both dyes suggesting accommodation of the common benzothiazole residue between base pairs. Experiments using poly(dA-dT)-poly(dA-dT) and poly(dG-dC)-poly(dG-dC) confirm base pair preference for TO. (c) 2007 Elsevier Inc. All rights reserved.
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