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Spectroscopic Characterization of the Interaction of Phenosafranin and Safranin O with Double Stranded, Heat Denatured and Single Stranded Calf Thymus DNA

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Abstract

Interaction of phenosafranin and safranin O with double stranded, heat denatured and single stranded calf thymus DNA has been studied by fluorescence, absorbance and circular dichroic techniques. Binding to the double stranded and heat denatured DNA conformations induced strong quenching in the fluorescence spectra of both dyes. Linear Scatchard plots indicated the binding to be of one type and the affinity evaluated to be of the order of 105 M−1 with double stranded and heat denatured DNAs. Fluorescence quenching was much weaker with the single stranded DNA and the binding affinity was one order lower. Ferrocyanide quenching studies revealed that the fluorescence emission of the dye molecules bound to the double stranded and heat denatured DNAs was quenched much less compared to that bound to the single stranded DNA. Further, there was significant emission polarization for the bound dyes and strong energy transfer from the DNA base pairs to the dye molecules indicating intercalative binding. Salt dependence of the binding phenomenon revealed that electrostatic forces have significant role in the binding process. The intercalation of these molecules to double stranded and heat denatured DNA and simple stacking to single strands was proved by these fluorescence techniques. Support to the fluorescence results have been derived from absorption and circular dichroic results. Phenosafranin was revealed to be a stronger binding species compared to safranin O.

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Acknowledgements

Ishita Saha is a National Eligibility Test qualified Junior Research Fellow of the Council of Scientific and Industrial Research (CSIR), Government of India. The authors thank Prof. Siddhartha Roy, Director, Indian Institute of Chemical Biology for his patronage and all colleagues of the Biophysical Chemistry Laboratory for their help and cooperation at every stage of this investigation.

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  1. Biophysical Chemistry Laboratory, Indian Institute of Chemical Biology, CSIR, 4, Raja S. C. Mullick Road, Kolkata, 700032, India

    Ishita Saha & Gopinatha Suresh Kumar

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  1. Ishita Saha
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  2. Gopinatha Suresh Kumar
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Correspondence to Gopinatha Suresh Kumar.

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Saha, I., Kumar, G.S. Spectroscopic Characterization of the Interaction of Phenosafranin and Safranin O with Double Stranded, Heat Denatured and Single Stranded Calf Thymus DNA. J Fluoresc 21, 247–255 (2011). https://doi.org/10.1007/s10895-010-0712-3

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  • DOI: https://doi.org/10.1007/s10895-010-0712-3

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