| Identification of nucleic acid high-affinity binding sequences of proteins by SELEX. | |
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MedLine Citation:
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PMID: 19378165 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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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Authors:
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Philippe Bouvet |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 543 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2009 |
Date Detail:
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Created Date: 2009-04-20 Completed Date: 2009-06-24 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 139-50 Citation Subset: IM |
Affiliation:
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Laboratoire de Biologie Moléculaire de la Cellule, CNRS UMR 5239, IFR128 Biosciences, 46 Allée d'Italie, Lyon, France. pbouvet@ens-lyon.fr |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Base Sequence Nucleic Acids / genetics* Protein Binding Proteins / metabolism* SELEX Aptamer Technique / methods* |
| Chemical | |
Reg. No./Substance:
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0/Nucleic Acids; 0/Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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