ABSTRACT During the last 25 y, fluorescent protein tagging has become a tool of choice to investigate protein function in a cellular context. The information gathered with this approach is not only providing insights into protein subcellular localization but also allows contextualizing protein function in multicellular settings. Here we illustrate the power of this method by commenting on the recent successful localization of the large membrane DEK1 protein during three-dimensional body formation in the moss Physcomitrella patens. But as many approaches, protein tagging is not exempt of caveats. The multiple infructuous (failed) attempts to detect DEK1 using a fluorescent protein tag present a good overview of such potential problems. Here we discuss the insertion of different fluorescent proteins at different positions in the PpDEK1 protein and the resulting unintended range of mutant phenotypes. Albeit none of these mutants generated a detectable fluorescent signal they can still provide interesting biological information about DEK1 function.

中文翻译：

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通过 DEK1 蛋白质透镜看到的植物体内蛋白质定位的挑战

摘要 在过去的 25 年中，荧光蛋白标记已成为研究细胞环境中蛋白质功能的首选工具。用这种方法收集的信息不仅提供了对蛋白质亚细胞定位的见解，而且还允许在多细胞环境中对蛋白质功能进行背景化。在这里，我们通过评论最近在苔藓 Physcomitrella patens 中的三维体形成过程中大膜 DEK1 蛋白的成功定位来说明这种方法的力量。但与许多方法一样，蛋白质标记也不能免除警告。使用荧光蛋白标签检测 DEK1 的多次徒劳（失败）尝试很好地概述了此类潜在问题。在这里，我们讨论在 PpDEK1 蛋白的不同位置插入不同的荧光蛋白以及由此产生的意外突变表型范围。尽管这些突变体都没有产生可检测的荧光信号，但它们仍然可以提供有关 DEK1 功能的有趣生物学信息。