Molecular and cell biology methods for fungi / edited by Amir Sharon. — New York : Humana Press, c2010. – (58.17/M592/v.638)

Contents

    Contents
    
    Contributors
    PART I FUNGAL TRANSFORMATION AND GENE KNOCKOUT
    1 Protoplast Transformation of Filamentous Fungi
    2 Electroporation and Agrobacterium-Mediated Spore Transformation
    3 High-Throughput Construction of Gene Deletion Cassettes for Generation of Neurospora crassa Knockout Strains
    4 Development of Impala-Based Transposon Systems for Gene Tagging in Filamentous Fungi
    5 DelsGate: A Robust and Rapid Method for Gene Deletion..
    6 Gene Silencing for Functional Analysis: Assessing RNAi as a Tool for Manipulation of Gene Expression
    PART II DETECTION AND QUANTIFICATION OF FUNGI
    7 Analysis of Fungal Gene Expression by Real Time Quantitative PCR
    8 Identification of Differentially Expressed Fungal Genes In Planta by Suppression Subtraction Hybridization
    9 Quantification of Fungal Infection of Leaves with Digital Images and Scion Image Software
    10 Expression Profiling of Fungal Genes During Arbuscular Mycorrhiza Symbiosis Establishment Using Direct Fluorescent In Situ RT-PCR.
    11 Application of Laser Microdissection to Study Plant-Fungal Pathogen Interactions 153
    12 Multiplex Gene Expression Analysis by TRAC in Fungal Cultures 165
    13 Amplification of Fungal Genomes Using Multiple Displacement Amplification
    PART III MICROSCOPY AND PROTEIN ANALYSIS
    14 Biochemical Methods Used to Study the Gene Expression and Protein Complexes in the Filamentous Fungus Neurospora crassa
    15 Measuring Protein Kinase and Sugar Kinase Activity in Plant Pathogenic Fusarium Species
    16 A Detailed Protocol for Chromatin Immunoprecipitation in the Yeast Saccharomyces cerevisiae
    17 A Method to Visualize the Actin and Microtubule Cytoskeleton by Indirect Immunofluorescence
    18 Fluorescence In Situ Hybridization for Molecular Cytogenetic Analysis in Filamentous Fungi
    19 Live-Cell Imaging of Microtubule Dynamics in Hyphae of Neurospora crassa
    20 Methods to Detect Apoptotic-Like Cell Death in Filamentous Fungi 269
    21 Evaluation of Antifungal Susceptibility Using Flow Cytometry
    22 Preparation of Fungi for Ultrastructural Investigations and Immunogoldlabelling
    23 Split-EGFP Screens for the Detection and Localisation of Protein-Protein Interactions in Living Yeast Cells
    Index