I.K. International Pvt. Ltd

Publication Year 2004

ISBN 9780195670837

ISBN-10 0195670833


Language (English)
New design Covers a multi-disciplinary approach to biological problems Vital for the study of macromolecular DNA-protein complexes Over 70 protocols Well illustrated with some colour figures DNA-Protein Interactions is a novel compilation of methods for studying the interactions of proteins with DNA. It is a rapidly advancing research area in which multidisciplinary approaches are especially valuable for solving problems and obtaining a detailed understanding of the molecular regulatory interactions involved. This book covers all the major tools that are required for the study of the large macromolecular enzymatic machines that manipulate DNA, with particular emphasis on biophysical techniques applied to the analysis of transcription and its relation to chromatin structure. Knowledge of basic techniques is assumed, although advances in fundamental fields are covered. Table Of Contents: Expression Systems Gel electrophoresis and bending in cisplatin-modified linear DNA Use of DNA microcircles in protein-DNA binding studies Use of topology to measure protein-induced DNA bend and unwinding angles Bandshift, gel retardation, or electrophoretic mobility shift assays Practical aspects of Fluorescence Resonance Energy Transfer (FRET) and its applications in nucleic acid biochemistry Determination of DNA-ligand interactions by fluorescence correlation spectroscopy DNA wrapping in E. Coli RNA polymerase open promotor complexes revealed by scanning force microscopy Microcalorimetry of protein-DNA interactions DNA-protein cross-linking by formaldehyde: a probe for RNAP-open promoter complex structure Solid-phase DNAse I footprinting Hydroxyl radical footprinting Radiolytic cleavage of DNA: Mapping of the Protein Interaction Sites UV - Laser photoreactivity of nucleoprotein complexes in vitro In vivo UV-laser footprinting Digitization and quantitative analysis of footprinting gels Mapping histone positions in chromatin by protein-directed DNA cross-linking and cleavage Kinetic