FRET-based analysis of protein-nucleic acid interactions by genetically incorporating a fluorescent amino acid.
Amino Acids
; 47(4): 729-34, 2015 Apr.
Article
em En
| MEDLINE
| ID: mdl-25540052
ABSTRACT
Protein-nucleic acid interaction is an important process in many biological phenomena. In this study, a fluorescence resonance energy transfer (FRET)-based protein-DNA binding assay has been developed, in which a fluorescent amino acid is genetically incorporated into a DNA-binding protein. A coumarin-containing amino acid was incorporated into a DNA-binding protein, and the mutant protein specifically produced a FRET signal upon binding to its cognate DNA labeled with a fluorophore. The protein-DNA binding affinity was then measured under equilibrium conditions. This method is advantageous for studying protein-nucleic acid interactions, because it is performed under equilibrium conditions, technically easy, and applicable to any nucleic acid-binding protein.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
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Transferência Ressonante de Energia de Fluorescência
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Proteínas de Ligação a DNA
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Aminoácidos
Tipo de estudo:
Evaluation_studies
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article