Exonuclease III footprinting on immobilized DNA templates.
DNA footprinting is a widely used method to locate the binding sites of protein on the DNA. It is based on the observation that a protein bound to DNA protects it from degradation by an enzyme or chemical reagent.Exonuclease III is a suitable probe to analyze the boundaries of a protein when it is necessary to eliminate any excess unbound DNA from the reaction to avoid background problems. In combination with biotin-labeled DNA that is bound to streptavidin-coated magnetic particles, information on the precise position of a DNA bound protein is available within a few hours. The position of the archaeal RNA polymerase at different stages of transcription in the Pyrococcus furiosus in vitro transcription system was analyzed by this method.
Spitalny
Patrizia
P
Department of Microbiology, University of Regensburg, Universitätsstrasse 31, 93053, Regensburg, Germany.
Thomm
Michael
M
D004247:DNA / Q000378:metabolism*; D018983:DNA Footprinting / Q000379:methods*; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D007553:Isotope Labeling; D020027:Pyrococcus furiosus / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III