Quantitative distance information on protein-DNA complexes determined in polyacrylamide gels by fluorescence resonance energy transfer.
作者： Mike Lorenz , Stephan Diekmann
DOI: 10.1002/1522-2683()22:6<990::AID-ELPS990>3.0.CO;2-X
关键词:
摘要: In polyacrylamide gels, we have quantitatively determined Forster transfer (fluorescense resonance energy transfer, FRET) between two fluorescent dyes attached to DNA in protein-DNA complexes. The donor-dye fluorescein labeled retains its free mobility the gel, however, fluorescence properties change. different quantum yield of gel is found be independent concentration and can thus taken into account by a reduced distance R0 46 A compared 50 solution. We global structural proteins binding double-labeled using novel gel-based assay. gels analyzed integration host factor (IHF) high group protein NHP6a their substrate DNA. measured efficiency allows us deduce information on overall shape bending angle complex.
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