title: Detecting DNA-binding of proteins in vivo by UV-crosslinking and immunoprecipitation.
abstract: The temporal and spatial binding of proteins on DNA is important to the regulation of genome expression and maintenance. However, examining how the protein-DNA complexes assemble in living cells is challenging. The development of UV-crosslinking/immunoprecipitation (UV-X-ChIP) technique and the progress of its applications show the powerful potential of this method in detecting such binding behavior in vivo. UV light is a zero length crosslinker and is believed to produce less perturbation of the complex than chemical crosslinker. The use of UV laser as UV light source allows the number of photons required for crosslinking to be delivered in nano- or pico- or femtosecond intervals, extremely shortening the irradiation time and achieving higher crosslinking efficiency than conventional UV lamp, thus being well suitable for kinetic studies. UV-X-ChIP technique has been successfully applied on the study of DNA replication, transcription, chromatin structure, and genome-wide location of DNA-binding proteins.
authors.affiliation: Laboratory for Conservation and Utilization of Bio-resource, Yunnan University, Kunming, Yunnan 650091, PR China. zhanglm@ynu.edu.cn
authors.forename: Lemin
authors.initials: L
authors.lastname: Zhang
authors.forename: Keqin
authors.initials: K
authors.lastname: Zhang
authors.forename: Ralf
authors.initials: R
authors.lastname: Prändl
authors.forename: Fritz
authors.initials: F
authors.lastname: Schöffl
mesh_terms: D000818:Animals; D001665:Binding Sites; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000528:radiation effects; D006801:Humans; D014466:Ultraviolet Rays
chemical_list: D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins