Summary
A technique is described for the identification of nucleic acid sequences bound with high affinity by proteins or by other molecules suitable for a partitioning assay. Here, a histidine-tagged protein is allowed to interact with a pool of nucleic acids and the protein–nucleic acid complexes formed are retained on a Ni-NTA matrix. Nucleic acids with a low level of recognition by the protein are washed away. The pool of recovered nucleic acids is amplified by the polymerase chain reaction and is submitted to further rounds of selection. Each round of selection increases the proportion of sequences that are avidly bound by the protein of interest. The cloning and sequencing of these sequences finally completes their identification.
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Acknowledgments
The work in the authors’ laboratory is supported by grants from the CNRS, ANR N° BLAN07-2_190263, and Association pour la Recherche sur le Cancer (ARC).
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Bouvet, P. (2009). Identification of Nucleic Acid High-Affinity Binding Sequences of Proteins by SELEX. In: Leblanc, B., Moss, T. (eds) DNA-Protein Interactions. Methods in Molecular Biology™, vol 543. Humana Press. https://doi.org/10.1007/978-1-60327-015-1_11
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DOI: https://doi.org/10.1007/978-1-60327-015-1_11
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