Abstract
We developed a technique of differential electrophoretic mobility shift assay (EMSA) display allowing identification of tissue-specific protein-binding sites within long genomic sequences. Using this approach, we identified 10 cell type-specific protein-binding sites (protein target sites [PTSs]) within a 137-kb human chromosome 19 region. In general, tissue-specific binding of proteins from different nuclear extracts by individual PTSs did not follow the all-or-nothing principle. Most often, PTS-protein complexes were formed in all cases, but they were different for different nuclear extracts used.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Base Sequence
-
Binding Sites / genetics
-
Cell Nucleus / metabolism
-
Cells, Cultured
-
Chromosome Mapping / methods*
-
DNA-Binding Proteins / chemistry*
-
Data Display*
-
Electrophoresis, Agar Gel
-
Electrophoresis, Gel, Two-Dimensional
-
Electrophoresis, Polyacrylamide Gel
-
Electrophoretic Mobility Shift Assay / instrumentation
-
Electrophoretic Mobility Shift Assay / methods*
-
Gene Library*
-
Genetic Techniques
-
Humans
-
Isoelectric Focusing
-
Nuclear Proteins / genetics
-
Sensitivity and Specificity
-
Tissue Distribution
-
Transcription Factors / chemistry*
Substances
-
DNA-Binding Proteins
-
Nuclear Proteins
-
Transcription Factors