Biochemical techniques for the characterization of G-quadruplex structures: EMSA, DMS footprinting, and DNA polymerase stop assay.
作者： Daekyu Sun , Laurence H. Hurley
DOI: 10.1007/978-1-59745-363-9_5
关键词: Polypyrimidine tract 、 G-quadruplex 、 DNA binding site 、 Biochemistry 、 DNA footprinting 、 Footprinting 、 Biology 、 DNase footprinting assay 、 Electrophoretic mobility shift assay 、 DNA clamp
摘要: The proximal promoter region of many human growth-related genes contains a polypurine/polypyrimidine tract that serves as multiple binding site for Sp1 or other transcription factors. These tracts often contain guanine-rich sequence consisting four runs three more contiguous guanines separated by one bases, corresponding to general motif known the formation an intramolecular G-quadruplex. Recent results provide strong evidence specific G-quadruplex structures can be formed naturally G-rich within these regions, raising possibility transcriptional control modulated G-quadruplex-interactive agents. In this chapter, we describe biochemical methodologies, electrophoretic mobility shift assay (EMSA), dimethylsulfate (DMS) footprinting, and DNA polymerase stop assay, which useful initial characterization sequences.
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