Devos Jonas, Van Dijck Patrick, Van Genechten Wouter
Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, Leuven, Belgium.
Bio Protoc. 2024 Oct 5;14(19):e5082. doi: 10.21769/BioProtoc.5082.
is the most common human fungal pathogen, able to reside in a broad range of niches within the human body. Even though systemic infection is associated with high mortality, the fungus has historically received relatively little attention, resulting in a lack of optimized molecular and fluorescent tools. Over the last decade, some extra focus has been put on the optimization of fluorescent proteins (FPs) of C. albicans. However, as the FPs are GFP-type, they require an aerobic environment and a relatively long period to fully mature. Recently, we have shown the application of a novel type of fluorogen-based FP, with an improved version of fluorescence activating and absorption shifting tag (iFAST), in . Due to the dynamic relation between iFAST and its fluorogens, the system has the advantage of being reversible in terms of fluorescence. Furthermore, the combination of iFAST with different fluorogens results in different spectral and cellular properties, allowing customization of the system. Key features • Genetic integration and tagging with the iFAST tag in • Imaging and localization of a protein of interest tagged with iFAST. • Reversibility of fluorescence with iFAST.
是最常见的人类真菌病原体，能够在人体内广泛的生态位中生存。尽管全身感染与高死亡率相关，但这种真菌在历史上受到的关注相对较少，导致缺乏优化的分子和荧光工具。在过去十年中，人们对白色念珠菌荧光蛋白（FPs）的优化给予了一些额外关注。然而，由于这些FPs是绿色荧光蛋白（GFP）类型，它们需要有氧环境和相对较长的时间才能完全成熟。最近，我们展示了一种新型的基于荧光原的FPs的应用，其具有改进版的荧光激活和吸收转移标签（iFAST），在……中。由于iFAST与其荧光原之间的动态关系，该系统在荧光方面具有可逆的优势。此外，iFAST与不同荧光原的组合会产生不同的光谱和细胞特性，从而可以对该系统进行定制。关键特性 • 在……中用iFAST标签进行基因整合和标记 • 对用iFAST标记的感兴趣蛋白质进行成像和定位 • iFAST荧光的可逆性
