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Analysis of Virus-Induced Double-Stranded RNA in Living Plant Cells by the dRBFC Assay
Ying Zhang1, Xinyue Fan1, Xiaofei Cheng2
1College of Plant Protection, Northeast Agricultural University, Harbin, Heilongjiang, China.
Abstract:
Double-stranded RNA (dsRNA) is the replicate intermediate of all RNA viruses, and is also recognized by their host cells as a virus-invading molecule signal. Analysis of the localization and dynamic of virus-induced dsRNA can reveal crucial information concerning the molecular mechanism of virus replication and host responses to viral infection. In this chapter, we provide an easy and efficient protocol called dsRNA binding-dependent fluorescence complementation (dRBFC) assay for labeling the dsRNAs in living plant cells using two different plant RNA viruses, namely potato virus X and turnip mosaic virus. Moreover, both YFP- and mRFP-based dRBFC plasmids are available for the flexibility of experiment design.
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