Identifying protein-protein interactions (PPI) is crucial for understanding biological processes. Many PPI tools are available, yet only some function within the context of a plant cell. Narrowing down even further, only a few tools allow complex multiprotein interactions to be visualized. Here, we present a conditional in vivo PPI tool for plant research that meets these criteria. Knocksideways in plants (KSP) is based on the ability of rapamycin to alter the localization of a bait protein and its interactors via the heterodimerization of FKBP and FRB domains. KSP is inherently free from many limitations of other PPI systems. This in vivo tool does not require spatial proximity of the bait and prey fluorophores and it is compatible with a broad range of fluorophores. KSP is also a conditional tool and therefore the visualization of the proteins in the absence of rapamycin acts as an internal control. We used KSP to confirm previously identified interactions in Nicotiana benthamiana leaf epidermal cells. Furthermore, the scripts that we generated allow the interactions to be quantified at high throughput. Finally, we demonstrate that KSP can easily be used to visualize complex multi-protein interactions. KSP is therefore a versatile tool with unique characteristics and applications that complements other plant PPI methods.

中文翻译：

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通过雷帕霉素依赖性离域可视化植物中的蛋白质相互作用

鉴定蛋白质相互作用（PPI）对于理解生物学过程至关重要。有许多PPI工具可用，但在工厂单元的环境中只有某些功能。进一步缩小范围，只有很少的工具可以使复杂的多蛋白相互作用可视化。在这里，我们提出了满足这些条件的用于植物研究的条件体内PPI工具。植物中的敲除现象（KSP）基于雷帕霉素通过FKBP和FRB结构域的异二聚化改变诱饵蛋白及其相互作用物的定位的能力。KSP本质上不受其他PPI系统的许多限制。这种体内工具不需要诱饵和猎物荧光团在空间上接近，并且与各种荧光团兼容。KSP还是有条件的工具，因此在不存在雷帕霉素的情况下可视化蛋白质可作为内部对照。我们使用KSP来确认先前确定的本生烟草叶表皮细胞中的相互作用。此外，我们生成的脚本允许以高吞吐量量化交互。最后，我们证明了KSP可以轻松地用于可视化复杂的多蛋白相互作用。因此，KSP是具有独特特性和应用的通用工具，可补充其他工厂PPI方法。我们证明KSP可以轻松地用于可视化复杂的多蛋白相互作用。因此，KSP是具有独特特性和应用的通用工具，可补充其他工厂PPI方法。我们证明KSP可以轻松地用于可视化复杂的多蛋白相互作用。因此，KSP是一种多功能工具，具有独特的特性和应用，可补充其他工厂PPI方法。