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A new electron microscopic method for studying protein-nucleic acid interactions.
Authors:M Zollinger  M Guertin  M D Mamet-Bratley
Affiliation:Département de Biochimie, Faculté de Médecine, Université de Montreal, Montreal, Québec H3C 3J7, Canada
Abstract:
A new method for preparation of nucleic acid specimens for electron microscopy has been adapted to study the interaction of proteins with DNA. Both a detergent and a basic protein are added to the DNA-protein solution before spreading on a hypophase containing 0.2 m ammonium acetate. This method has been tested using T7 DNA and Escherichia coli RNA polymerase. Specifically bound enzyme molecules were clearly visible on the well extended DNA molecules; the binding sites were located at 0.59, 1.24, 1.57, and 1.86% of the total length of T7 DNA. Under carefully controlled conditions, 40–85% of the DNA molecules specifically bound at least one enzyme molecule.
Keywords:
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