Fungi

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    Protocols in Current Issue
    High Resolution Respirometry in Candida albicans
    Authors:  Lucian Duvenage, Carol A. Munro and Campbell W. Gourlay, date: 09/05/2019, view: 141, Q&A: 0
    [Abstract] Many Candida species, such as the opportunistic human pathogen Candida albicans, are Crabtree-Negative yeasts and are therefore highly dependent on the energy generated through oxidative phosphorylation. Respiration control is linked to a range of aspects of C. albicans cell physiology that appear to be important for ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 235, Q&A: 0
    [Abstract] In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
    Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence
    Authors:  José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 426, Q&A: 0
    [Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...
    Method for CRISPR/Cas9 Mutagenesis in Candida albicans
    Authors:  Neta Dean and Henry Ng, date: 04/20/2018, view: 4489, 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: 3496, 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: 4382, 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: 4618, 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 ...
    Large-scale Phenotypic Profiling of Gene Deletion Mutants in Candida glabrata
    Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 6889, Q&A: 0
    [Abstract] Here, we describe a method enabling the phenotypic profiling of genome-scale deletion collections of fungal mutants to detect phenotypes for various stress conditions. These stress conditions include among many others antifungal drug susceptibility, temperature-induced and osmotic as well as heavy metal or oxidative stress. The protocol was ...
    Genetic Transformation of Candida glabrata by Heat Shock
    Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 5963, Q&A: 0
    [Abstract] Here, we report a method for the transformation of Candida glabrata using a heat shock method. The protocol can be used for transformations in single well or in 96-well scale. It has been employed as an alternative method to the electroporation protocol to construct a genome-scale gene deletion collection in the human fungal pathogen ...
    Genetic Transformation of Candida glabrata by Electroporation
    Authors:  Fabian Istel, Tobias Schwarzmüller, Michael Tscherner and Karl Kuchler, date: 07/20/2015, view: 5318, Q&A: 0
    [Abstract] Here, we report a method for the transformation by electroporation of the human fungal pathogen Candida glabrata (C. glabrata). The protocol can be used for transformations in single well or in 96-well microtiter plates. It has been extensively used to generate a genome-scale gene deletion library using the C. glabrata ...



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