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 ...
    In vitro Deneddylation Assay
    Authors:  Anna M. Köhler, Cindy Meister and Gerhard H. Braus, date: 03/20/2016, view: 8336, 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) ...
    XTT Assay of Antifungal Activity
    Authors:  Flávio V. Loures and Stuart M. Levitz, date: 08/05/2015, view: 9098, 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: 7664, 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 ...
    Western Analysis of Histone Modifications (Aspergillus nidulans)
    Authors:  Alexandra Soukup and Nancy P. Keller, date: 04/05/2013, view: 11132, Q&A: 0
    [Abstract] Western blotting allows for the specific detection of proteins and/or modifications of proteins by an antibody of interest. This protocol utilizes a crude nuclei extraction protocol for Aspergillus nidulans to enrich for histones and other nuclear proteins prior to gel electrophoresis. Post translational modifications of histones may ...



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