Characterization of DNA-binding proteins using multiplexed competitor EMSA.
We describe a low-cost high-throughput technique to characterize nuclear protein DNA-binding interactions. This technique, known as Multiplexed Competitor Electrophoretic Mobility Shift Assay, uses a series of multiplexed oligonucleotide DNA consensus competitors, in combination with a standard electrophoretic mobility shift assay procedure, to efficiently characterize DNA-binding proteins. We show utility for the method to identify a previously unreported hepatocyte nuclear factor-3 site created in intron 8 of the lipoprotein lipase gene by a common single-nucleotide polymorphism (rs327).
Smith
Andrew J P
AJ
Center for Cardiovascular Genetics, Department of Medicine, University College London, 5 University Street, London WC1E 6JF, UK. Andrew.J.P.Smith@ucl.ac.uk.
Humphries
Steve E
SE
D001483:Base Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*
D017931:DNA Primers; D004268:DNA-Binding Proteins