Determination of Adeno-associated Virus Rep DNA Binding Using Fluorescence Anisotropy

Francisco Zarate-Perez, Vishaka Santosh, Martino Bardelli, Leticia Agundez, R. Michael Linden, Els Henckaerts, Carlos Escalante
2017 Bio-protocol  
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.
doi:10.21769/bioprotoc.2194 fatcat:s64yhdyeajbbbnnvtvc64pykxa