Document Detail


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

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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