Determination of Adeno-associated Virus Rep DNA Binding Using Fluorescence Anisotropy
نویسندگان
چکیده
منابع مشابه
Determination of Adeno-associated Virus Rep DNA Binding Using Fluorescence Anisotropy.
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...
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Both the Rep68 and Rep78 proteins of adeno-associated virus type 2 (AAV) bind to AAV terminal repeat hairpin DNA and can mediate site-specific nicking in vitro at the terminal resolution site (trs) within the terminal repeats. To define the regions of the Rep proteins required for these functions, a series of truncated Rep78 derivatives was created. Wild-type and mutant proteins were synthesize...
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Integration into a particular location in human chromosomes is a unique property of the adeno-associated virus (AAV). This reaction requires the viral Rep protein and AAV origin sequences. To understand how Rep recognizes DNA, we have determined the structures of the Rep endonuclease domain separately complexed with two DNA substrates: the Rep binding site within the viral inverted terminal rep...
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ژورنال
عنوان ژورنال: BIO-PROTOCOL
سال: 2017
ISSN: 2331-8325
DOI: 10.21769/bioprotoc.2194