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DNA–WT1 protein interaction studied by surface-enhanced Raman spectroscopy

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Abstract

Interactions of proteins with DNA play an important role in regulating the biological functions of DNA. Here we propose and demonstrate the detection of protein–DNA binding using surface-enhanced Raman scattering (SERS). In this method, double-stranded DNA molecules with potential protein-binding sites are labeled with dye molecules and immobilized on metal nanoparticles. The binding of proteins protects the DNA from complete digestion by exonuclease and can be detected by measuring the SERS signals before and after the exonuclease digestion. As a proof of concept, this SERS-based protein–DNA interaction assay is validated by studying the binding of a zinc finger transcription factor WT1 with DNA sequences derived from the promoter of the human vascular endothelial growth factor.

A representative dark-field optical microscopic image of Ag nanoparticles attached with dye labeled DNA molecules and two Raman spectra from AAg nanoparticles attached with FAM (left) and TAMRA (right) labeled DNA molecules respectively.

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Acknowledgments

We acknowledge the financial support from the Ohio Board of Regents through a research challenge grant. H. A. is supported by an NSF REU program through a grant CHE0649017.

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Authors and Affiliations

  1. Liquid Crystal Institute, Kent State University, Kent, OH, 44242, USA

    Bhuwan Joshi, Ayan Chakrabarty, Christopher Bruot, Hannah Ainsworth & Qi-Huo Wei

  2. Department of Biological Science, Kent State University, Kent, OH, 44242, USA

    Gail Fraizer

Authors
  1. Bhuwan Joshi
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  2. Ayan Chakrabarty
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  3. Christopher Bruot
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  4. Hannah Ainsworth
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  5. Gail Fraizer
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  6. Qi-Huo Wei
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Corresponding authors

Correspondence to Gail Fraizer or Qi-Huo Wei.

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Joshi, B., Chakrabarty, A., Bruot, C. et al. DNA–WT1 protein interaction studied by surface-enhanced Raman spectroscopy. Anal Bioanal Chem 396, 1415–1421 (2010). https://doi.org/10.1007/s00216-009-3364-x

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  • DOI: https://doi.org/10.1007/s00216-009-3364-x

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