Detection of protein-DNA interaction with a DNA probe: distinction between single-strand and double-strand DNA-protein interaction.
Nucleic Acids Res
; 32(13): e110, 2004 Jul 25.
Article
en En
| MEDLINE
| ID: mdl-15273279
A simple, direct method for the detection of DNA-protein interaction was developed with electrochemical methods. Single-stranded DNA (ss-DNA) probes were prepared through the chemical bonding of an oligonucleotide to a polymer film bearing carboxylic acid groups, and double-stranded DNA (ds-DNA) probes were prepared through hybridization of the complementary sequence DNA on the ss-DNA probe. Impedance spectroscopy and differential pulse voltammetry (DPV) distinguished the interaction between the DNA probes with mouse Purbeta (mPurbeta), an ss-DNA binding protein, and with Escherichia coli MutH, a ds-DNA binding protein. Impedance spectra obtained before and after the interaction of DNA probes with these proteins clearly showed the sequence-specific ss-DNA preference of mPurbeta and the sequence-specific ds-DNA preference of MutH. The concentration dependence of proteins on the response of the DNA probes was also investigated, and the detection limits of MutH and mPurbeta were 25 and 3 microg/ml, respectively. To confirm the impedance results, the variation of the current oxidation peak of adenine of the DNA probe was monitored with DPV. The formation constants of the complexes formed between the probe DNA and the proteins were estimated based on the DPV results.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ADN
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ADN de Cadena Simple
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Sondas de ADN
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Proteínas de Unión al ADN
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Electroquímica
Tipo de estudio:
Diagnostic_studies
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Evaluation_studies
Idioma:
En
Revista:
Nucleic Acids Res
Año:
2004
Tipo del documento:
Article
Pais de publicación:
Reino Unido