Fungi

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    Protocols in Current Issue
    A Reliable Assay to Evaluate the Virulence of Aspergillus nidulans Using the Alternative Animal Model Galleria mellonella (Lepidoptera)
    [Abstract] The greater wax moth Galleria mellonella has emerged as an effective heterologous host to study fungal pathogenesis and the efficacy of promising antifungal drugs (Mylonakis et al., 2005; Li et al., 2013). Here, a methodology describing the Aspergillus nidulans infection in G. mellonella larvae, along ...
    Flow Cytometry of Lung and Bronchoalveolar Lavage Fluid Cells from Mice Challenged with Fluorescent Aspergillus Reporter (FLARE) Conidia
    Authors:  Anupam Jhingran, Shinji Kasahara and Tobias M Hohl, date: 09/20/2016, view: 8595, Q&A: 0
    [Abstract] Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding ...
    An Improved and Simplified Radial Gel Diffusion Assay for the Detection of Xylanase Activity
    Authors:  Raviraj M. Kalunke, Ilaria Moscetti, Silvio Tundo and Renato D’Ovidio, date: 07/05/2016, view: 5286, Q&A: 0
    [Abstract] Xylanase (E.C. 3.2.1.8) degrades β-1, 4 xylan by cleaving β-1, 4 glycosidic linkages randomly, resulting in the generation of xylose and xylo-oligosaccharides. Xylanases are produced by organisms including fungi, bacteria, yeast, marine algae, protozoans, snails, crustaceans and insects. Xylanases present considerable industrial interest for their ...
    Aspergillus terreus Infection of Fruits and Terrein Quantification by HPLC Analysis
    Authors:  Markus Gressler and Matthias Brock, date: 06/20/2016, view: 4513, Q&A: 0
    [Abstract] The opportunistic fungal human and plant pathogen Aspergillus terreus (A. terreus) can be isolated from sea water, soil or decaying organic matter such as rotting leaves and fruits. While growing on fruits A. terreus produces secondary metabolites such as terrein, which may ease its penetration into plant tissues. In ...
    In vitro Deneddylation Assay
    Authors:  Anna M. Köhler, Cindy Meister and Gerhard H. Braus, date: 03/20/2016, view: 5317, Q&A: 0
    [Abstract] Nedd8 is a small ubiquitin-like protein (9 kDa) covalently attached to a conserved lysine residue of a cullin protein which is part of cullin-RING ligases (CRLs). CRLs are major E3 ligases important for protein ubiquitination in the ubiquitin-proteasome pathway (UPP). The activity of CRLs is regulated by cycles of neddylation (CulA-N8, ~98 kDa) ...
    Establishing a Biofilm Co-culture of Pseudomonas and Aspergillus for Metabolite Extraction
    Authors:  He Zheng, Nancy P. Keller and Yun Wang, date: 12/05/2015, view: 7057, Q&A: 0
    [Abstract] Filamentous fungi and bacteria form mixed-species biofilms in nature and diverse clinical contexts (Frey-Klett et al., 2011; Peleg et al., 2010). The interactions between fungi and bacteria, often mediated by secreted metabolites, have important ramifications for the biology of the interacting partners (Frey-Klett et al. ...
    XTT Assay of Antifungal Activity
    Authors:  Flávio V. Loures and Stuart M. Levitz, date: 08/05/2015, view: 6115, Q&A: 0
    [Abstract] XTT assay is a colorimetric method that uses the tetrazolium dye, 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide (XTT) to quantify cell-mediated damage to fungi. Actively respiring fungal cells convert the water-soluble XTT to a water-soluble, orange colored formazan product (Meshulam et al., 1995). Here, we ...
    Purification of the GfsA-3x FLAG Protein Expressed in Aspergillus nidulans
    [Abstract] GfsA is a fungal β-galactofuranosyltransferase involved in the biosynthesis of O-glycan. To investigate the enzymatic functions of GfsA, we attempted to obtain a recombinant protein of this enzyme from two heterologous host organisms. However, GfsA could not be expressed as a recombinant protein in either Escherichia coli (E. ...
    Determination of D-galactofuranose Content of Galactomannoproteins in Aspergillus nidulans
    [Abstract] Galactofuranose (Galf) is a component of several polysaccharides and glycoconjugates in certain species of filamentous fungi. Galf residues are frequently found in Aspergillus glycoproteins, including N-glycans and O-mannose glycans that modify many cell wall proteins and extracellular enzymes. It is ...
    Transport Assays in Aspergillus nidulans
    Authors:  Emilia Krypotou and George Diallinas, date: 11/20/2013, view: 5478, Q&A: 0
    [Abstract] Transport assays allow the direct kinetic analysis of a specific transporter by measuring apparent Km and Vmax values, and permit the characterization of substrate specificity profiles through competition assays. In this protocol, we describe a rapid and easy method for performing uptake assays in the model ...



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