Abstract:
:Protein-binding DNA sequence elements encode a variety of regulated functions of genomes. Information about such elements is currently in a state of rapid growth, but improved methods are required to characterize the sequence specificity of DNA-binding proteins. We have established an in vitro method for specific and sensitive solution-phase analysis of interactions between proteins and nucleic acids in nuclear extracts, based on the proximity ligation assay. The reagent consumption is very low, and the excellent sensitivity of the assay enables analysis of as few as 1-10 cells. We show that our results are highly reproducible, quantitative, and in good agreement with both EMSA and predictions obtained by using a motif finding software. This assay can be a valuable tool to characterize in-depth the sequence specificity of DNA-binding proteins and to evaluate effects of polymorphisms in known transcription factor binding sites.
journal_name
Proc Natl Acad Sci U S Aauthors
Gustafsdottir SM,Schlingemann J,Rada-Iglesias A,Schallmeiner E,Kamali-Moghaddam M,Wadelius C,Landegren Udoi
10.1073/pnas.0611229104subject
Has Abstractpub_date
2007-02-27 00:00:00pages
3067-72issue
9eissn
0027-8424issn
1091-6490pii
0611229104journal_volume
104pub_type
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