利用 dRBFC 分析法分析活体细胞中的病毒诱导双链 RNA。

Analysis of Virus-Induced Double-Stranded RNA in Living Plant Cells by the dRBFC Assay.

机构信息

College of Plant Protection, Northeast Agricultural University, Harbin, Heilongjiang, China.

出版信息

Methods Mol Biol. 2024;2771:27-33. doi: 10.1007/978-1-0716-3702-9_5.

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.

摘要

双链 RNA（dsRNA）是所有 RNA 病毒的复制中间体，也被宿主细胞识别为病毒入侵分子信号。分析病毒诱导的 dsRNA 的定位和动态可以揭示病毒复制的分子机制和宿主对病毒感染的反应的关键信息。在本章中，我们提供了一种简单有效的方法，称为 dsRNA 结合依赖性荧光互补（dRBFC）测定法，用于标记马铃薯病毒 X 和芜菁花叶病毒这两种不同的植物 RNA 病毒在活植物细胞中的 dsRNA。此外，还提供了基于 YFP 和 mRFP 的 dRBFC 质粒，以满足实验设计的灵活性。