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Nucleic Acid-Binding Assay of Argonaute Protein Using Fluorescence Polarization

  • Tomohiro MiyoshiEmail author
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Part of the Methods in Molecular Biology book series (MIMB, volume 1680)

Abstract

Nucleic acid binding by the Argonaute protein is an important trigger step in the Argonaute-dependent gene silencing system. We established an in vitro method to detect the nucleic acid binding activity of the Argonaute protein by fluorescence polarization. In this chapter, we will describe the expression and purification of the prokaryotic (Rhodobacter sphaeroides) Argonaute protein, and the nucleic acid-binding analysis using a Fluorescence Polarization System (Beacon 2000).

Key words

Fluorescence polarization assay Beacon 2000 Rhodobacter sphaeroides argonaute Guide RNA Purification of prokaryotic argonaute 

Notes

Acknowledgments

This work was supported by the JSPS KAKENHI (Grant Numbers: 25840019 and 16K07246), the Takeda Science Foundation, the Naito Foundation, the Sasaki Environment Technology Foundation, and the Union Tool Scholarship Foundation.

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© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Center for Transdisciplinary ResearchNiigata UniversityNiigataJapan

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