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首页> 外文期刊>Nucleus >Nucleolus-tethering system (NoTS) An intranuclear fluorescence two-hybrid assay for studying the nuclear protein-protein interactions and nuclear body initiation in vivo
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Nucleolus-tethering system (NoTS) An intranuclear fluorescence two-hybrid assay for studying the nuclear protein-protein interactions and nuclear body initiation in vivo

机译:核仁系留系统(NoTS)核内荧光双杂交测定法,用于研究体内核蛋白-蛋白相互作用和核体起始

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摘要

Multiple biological processes are regulated by complicated interaction networks formed by protein-protein or protein-RNA interactions. Nuclear bodies (NBs) are a class of membrane-less subnuclear structures, acting as reaction sites, storage and modification sites, or transcription regulating sites involved in signaling transduction. Biochemical and fluorescence-based methods are widely used to study protein-protein interactions, but false-positive results are a major issue, especially for some fluorescence-based methods. Moreover, these methods fail to be applied to study the formation of NBs, which were characterized by a popular bacterial Lac operator and/or repressor (LacO/LacI) system in mammalian cells. Methods investigating assembly of plant NBs are not available. We have recently developed a nucleolar marker protein nucleolin2 (Nuc2)-based method named Nucleolus-tethering System (NoTS) and showed its application in interaction assay among nucleoplasmic proteins and initiation of plant specific NBs, photobodies. In this extraview, we will compare NoTS with the traditional methods and discuss the assembly mechanisms of NBs, in addition to advantages, limitations, and perspectives about the application of NoTS.
机译:多种生物学过程受蛋白质-蛋白质或蛋白质-RNA相互作用形成的复杂相互作用网络调控。核小体(NBs)是一类无膜的亚核结构,充当信号转导中涉及的反应位点,存储和修饰位点或转录调控位点。生化和基于荧光的方法被广泛用于研究蛋白质之间的相互作用,但是假阳性结果是一个主要问题,尤其是对于某些基于荧光的方法。此外,这些方法未能用于研究NBs的形成,而NBs的特征是哺乳动物细胞中流行的细菌Lac操纵子和/或阻遏物(LacO / LacI)系统。无法使用调查植物NB组装的方法。我们最近开发了一种基于核仁标记蛋白nucleolin2(Nuc2)的方法,称为Nucleolus-tethering System(NoTS),并显示了其在核质蛋白之间相互作用测定和植物特异性NBs,光体起始中的应用。在此概述中,我们将比较NoTS与传统方法,并讨论NB的组装机制,以及关于NoTS应用的优点,局限性和观点。

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