Fungi

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    Protocols in Current Issue
    Method for CRISPR/Cas9 Mutagenesis in Candida albicans
    Authors:  Neta Dean and Henry Ng, date: 04/20/2018, view: 3712, Q&A: 0
    [Abstract] Candida albicans is the most prevalent and important human fungal pathogen. The advent of CRISPR as a means of gene editing has greatly facilitated genetic analysis in C. albicans. Here, we describe a detailed step-by-step procedure to construct and analyze C. albicans deletion mutants. This protocol uses plasmids that ...
    A Flow-assay for Farnesol Removal from Adherent Candida albicans Cultures
    Authors:  Melanie Polke and Ilse D. Jacobsen, date: 10/05/2017, view: 3065, Q&A: 0
    [Abstract] Here, we describe a protocol for a continuous flow system for C. albicans cultures growing adherent to a plastic surface. The protocol was adapted from a previous method established to simulate blood flow on endothelial cells (Wilson and Hube, 2010). The adapted protocol was used by us for the removal of molecules in C. albicans ...
    Fluorescently Labelled Aerolysin (FLAER) Labelling of Candida albicans Cells
    Authors:  Sneh Lata Singh and Sneha Sudha Komath, date: 06/05/2017, view: 3824, Q&A: 0
    [Abstract] In this protocol we describe a nonradiolabelled labelling of GPI anchor in Candida albicans. The method uses a fluorescent probe to bind specifically to GPI anchors so that the level of GPI-anchored proteins at the cell surface can be measured. The labelling does not need permeabilization of cells and can be carried out in vivo.
    In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae
    Authors:  Alex Hopke and Robert T. Wheeler, date: 04/05/2017, view: 4199, Q&A: 0
    [Abstract] In this protocol we describe how to visualize neutrophil extracellular traps (NETs) and fungal cell wall changes in the context of the coculture of mouse neutrophils with fungal hyphae of Candida albicans. These protocols are easily adjusted to test a wide array of hypotheses related to the impact of immune cells on fungi and the cell ...
    Detection of Hog1 Phosphorylation in Candida albicans in Response to an Antifungal Protein
    Authors:  Brigitte ME Hayes and Nicole L van der Weerden, date: 09/20/2014, view: 5652, Q&A: 0
    [Abstract] It is becoming increasingly apparent that stress signalling is important for tolerance of fungal species to antifungal chemicals and proteins. The high-osmolarity glycerol (HOG) pathway responds to a number of stressors including osmotic and oxidative stress. This protocol describes a method to detect activation of the Candida albicans ( ...
    ChIP-Seq in Candida albicans
    [Abstract] Systems biology approaches can be used to study the regulatory interactions occurring between many components of the biological system at the whole-genome level and decipher the circuitries implicated in the regulation of cellular processes, including those imparting virulence to opportunistic fungi. Candida albicans (C. albicans ...
    In vitro Analysis for Macrophage Binding and Pro-inflammatory Responses to Candida albicans
    Author:  Mohlopheni J. Marakalala, date: 05/05/2014, view: 5497, Q&A: 0
    [Abstract] Macrophage recognition of Candida albicans (C. albicans) is facilitated by pattern recognition receptors that interact with the fungal pathogen associated molecular patterns (PAMPs). Dectin-1 is the major macrophage receptor that is known to recognize fungal Beta-glucans leading to induction of various immune responses. This ...
    Immunoblot Analysis of Histone H4 Acetylation and Histone H2A Phosphorylation in Candida albicans
    Authors:  Michael Tscherner and Karl Kuchler, date: 10/20/2013, view: 6099, Q&A: 0
    [Abstract] Posttranslational modifications of histones are required for different processes including transcription, replication and DNA damage repair. This protocol describes the preparation of a whole-cell extracts for the fungal pathogen Candida albicans. Furthermore, the extract is used to detect lysine acetylation of histone H4 as well as ...
    Preparation of Candida albicans Biofilms for Transmission Electron Microscopy
    Authors:  Heather T. Taff and David R. Andes, date: 07/20/2013, view: 7387, Q&A: 0
    [Abstract] Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida albicans biofilms in ...
    Candida albicans Mitochondrial Protein Import Assay
    Authors:  Victoria L Hewitt, Trevor Lithgow and Ana Traven, date: 02/20/2013, view: 6480, Q&A: 0
    [Abstract] We have established the use of Candida albicans as a new model system to study mitochondrial biogenesis. This dimorphic yeast provides an excellent system to investigate the coordination of mitochondrial biogenesis with other cellular networks including cellular metabolism and the cell cycle. Unlike the model lab yeast Saccharomyces ...



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