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Quantitative measurement of proteins binding to DNA is a requisite to fully characterize the structural determinants of complex formation necessary to understand the DNA transactions that regulate cellular processes. Here we describe a detailed protocol to measure binding affinity of the adeno-associated virus (AAV) Rep68 protein for the integration site AAVS1 using fluorescent anisotropy. This protocol can be used to measure the binding constants of any DNA binding protein provided the substrate DNA is fluorescently labeled.

Type

Journal article

Journal

Bio-protocol

Publication Date

03/2017

Volume

7

Addresses

Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA.