Frazer Corey, Hernday Aaron D, Bennett Richard J
Molecular Microbiology and Immunology Department, Brown University, Providence, Rhode Island.
Department of Molecular and Cell Biology, School of Natural Sciences, University of California, Merced, Merced, California.
Curr Protoc Microbiol. 2019 Jun;53(1):e76. doi: 10.1002/cpmc.76. Epub 2019 Feb 12.
Candida albicans is an opportunistic human fungal pathogen that is able to cause both mucosal and systemic infections. It is also a frequent human commensal, where it is typically found inhabiting multiple niches including the gastrointestinal tract. One of the most remarkable features of C. albicans biology is its ability to undergo heritable and reversible switching between different phenotypic states, a phenomenon known as phenotypic switching. This is best exemplified by the white-opaque switch, in which cells undergo epigenetic transitions between two alternative cellular states. Here, we describe assays to quantify the frequency of switching between states, as well as methods to help identify cells in different phenotypic states. We also describe the use of environmental cues that can induce switching into either the white or opaque state. Finally, we introduce the use of mNeonGreen and mScarlet fluorescent proteins that have been optimized for use in C. albicans and which outperform commonly used fluorescent proteins for both fluorescence microscopy and flow cytometry. © 2019 by John Wiley & Sons, Inc.
白色念珠菌是一种机会性人类真菌病原体，能够引起黏膜感染和全身感染。它也是人类常见的共生菌，通常在包括胃肠道在内的多个生态位中被发现。白色念珠菌生物学最显著的特征之一是其能够在不同表型状态之间进行可遗传和可逆的转换，这一现象被称为表型转换。白色-不透明转换是最好的例证，在此过程中细胞在两种交替的细胞状态之间经历表观遗传转变。在这里，我们描述了量化状态间转换频率的测定方法，以及有助于识别不同表型状态细胞的方法。我们还描述了可诱导转换为白色或不透明状态的环境线索的使用。最后，我们介绍了mNeonGreen和mScarlet荧光蛋白的使用，它们已针对白色念珠菌进行了优化，在荧光显微镜和流式细胞术中均优于常用的荧光蛋白。© 2019约翰威立父子公司版权所有
