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Author:
Date:
2017-10-20
[Abstract] RNA-Protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA/protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-Protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-Protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) (Schonberger et al., 2012). Target RNA is tagged with 6xMS2 and MCP and RNA binding ...
[摘要] RNA-蛋白质相互作用在各种真核生物过程中起重要作用。 RNA /蛋白质复合物在植物中亚细胞定位的分子成像对于理解这些相互作用至关重要。然而，到目前为止，尚未开发在活植物中形成RNA-蛋白质相互作用的方法。最近，我们开发了一种三分子荧光互补（TriFC）系统，用于在烟草叶中瞬时表达的RNA-蛋白质相互作用的体内可视化。在这种方法中，我们将传统的双分子荧光互补（BiFC）系统与MS2系统（噬菌体MS2外壳蛋白[MCP]及其结合RNA序列[MS2序列]）（Schonberger等人，2012）相结合， 。目标RNA用6xMS2标记，MCP和RNA结合蛋白与YFP片段融合。编码这种融合RNA和蛋白质的DNA构建体用土壤杆菌悬浮液渗入烟草叶中。通过共焦显微镜观察体内的RNA-蛋白质相互作用
【背景】近来，多种类型的长非编码RNA（lncRNA）已经被鉴定并显示出在转录调节和染色质修饰中起重要作用（St Laurent等人，2015）。到目前为止，lncRNA介导的功能的大多数分子机制与RNA-蛋白质相互作用密切相关（St ...
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