《蛋白质-核酸相互作用技术》相关实验技术-Identification of tissue-specific DNA-protein binding sites by EMSA.pdfVIP

ANALYTICAL BIOCHEMISTRY Analytical Biochemistry 364 (2007) 60–66 /locate/yabio IdentiWcation of tissue-speciWc DNA–protein binding sites by means of two-dimensional electrophoretic mobility shift assay display Igor P. Chernov, Kira A. Timchenko, Sergey B. Akopov, Lev G. Nikolaev ¤, Eugene D. Sverdlov Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia Received 11 December 2006 Available online 2 February 2007 Abstract We developed a technique of diVerential electrophoretic mobility shift assay (EMSA) display allowing identiWcation of tissue-speciWc protein-binding sites within long genomic sequences. Using this approach, we identiWed 10 cell type-speciWc protein-binding sites (protein target sites [PTSs]) within a 137-kb human chromosome 19 region. In general, tissue-speciWc binding of proteins from diVerent 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 diVerent for diVerent nuclear extracts used. © 2007 Elsevier Inc. All rights reserved. Keywords: DNA–protein interactions; Electrophoretic mobility shift assay (EMSA); DiVerential display The publication of the draft human genome sequence Most of the gen