用 dRBFC 分析法分析植物活细胞中病毒诱导的双链 RNA。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2024-01-01 DOI:10.1007/978-1-0716-3702-9_5

Ying Zhang, Xinyue Fan, Xiaofei Cheng

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引用次数: 0

摘要

双链 RNA（dsRNA）是所有 RNA 病毒的复制中间体，也是宿主细胞识别的病毒入侵分子信号。对病毒诱导的dsRNA的定位和动态分析可以揭示病毒复制的分子机制和宿主对病毒感染的反应等重要信息。在本章中，我们利用两种不同的植物 RNA 病毒（即马铃薯病毒 X 和芜菁花叶病毒），提供了一种名为 dsRNA 结合依赖性荧光互补（dRBFC）的简便高效的方案，用于标记活体植物细胞中的 dsRNA。此外，还提供基于 YFP 和 mRFP 的 dRBFC 质粒，以便灵活设计实验。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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

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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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.