title: Identification of Nucleic Acid High Affinity Binding Sequences of Proteins by SELEX.
abstract: 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.
authors.affiliation: Ecole Normale Supérieure de Lyon, Laboratoire Joliot-Curie CNRS USR 3010, Université de Lyon, Lyon, France. pbouvet@ens-lyon.fr.|Laboratoire de Biologie Moléculaire de la Cellule, CNRS UMR 5239, IFR128 Biosciences, 46 Allée d'Italie, Lyon, France. pbouvet@ens-lyon.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bouvet
mesh_terms: D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D009696:Nucleic Acids / Q000737:chemistry / Q000235:genetics*; D052156:SELEX Aptamer Technique / Q000379:methods*
chemical_list: D004268:DNA-Binding Proteins; D009696:Nucleic Acids
keywords: Binding sequences; Nucleic acids ligands; SELEX; Sequence recognition