The Electrophoretic Mobility Shift Assay (EMSA) for Detection and Analysis of Protein-DNA Interactions

摘要: The interaction of proteins with specific DNA sites is a principal step in the control many cellular processes. Over past 20 years, our knowledge protein-DNA interactions has grown geometrically. This growth been facilitated by availability fast, reproducible assays for binding. Most popular these electrophoresis mobility-shift assay (EMSA), also known as “gel shift” or mobility assay, first described Garner and Revzin, Fried Crothers (Garner Revzin 1981; 1981). A particular strength this method its ability to detect simultaneous binding several single species, one (or more) protein(s) multiple species (Fried 1986; 1989), because DNA-complexes containing play central roles important transactions (e.g., replication, repair, transcription recombination) measurement allows assessment cooperative between within an assembly (Hudson 1990; Vossen et al. 1996). With minor variations, same techniques can be used follow rates decay protein-nucleic acid complexes (reviewed Gerstle 1993).