Click to have a closer look
About this book
About this book
With the development of fungal transformation systems and the deciphering of an increasing number of fungal genomes, this diverse clade of heterotrophic eukaryotic organisms has proven to be ideal for molecular work and highly serviceable as model systems to study basic processes with results that are applicable to many organisms, including humans.
In this volume, experts in the field provide an up-to-date set of practical protocols covering a range of frequently used methods used to study molecular and cellular aspects of fungal biology.
Part I: Fungal Transformation and Gene Knockout 1. Protoplast Transformation of Filamentous Fungi Gillian B. Turgeon, Bradford Condon, Jinyuan Liu, and Ning Zhang 2. Electroporation and Agrobacterium-Mediated Spore Transformation Anna Minz and Amir Sharon 3. High-Throughput Construction of Gene Deletion Cassettes for Generation of Neurospora crassa Knockout Strains Patrick D. Collopy, Hildur V. Colot, Gyungsoon Park, Carol Ringelberg, Christopher M. Crew, Katherine A. Borkovich, and Jay C. Dunlap 4. Development of Impala-Based Transposon Systems for Gene Tagging in Filamentous Fungi Marie Dufresne and Marie-Josee Daboussi 5. DelsGate: A Robust and Rapid Method for Gene Deletion Maria D. Garcia-Pedrajas, Marina Nadal, Timothy Denny, Lourdes Baeza-Montanez, Zahi Paz, and Scott E. Gold 6. Gene Silencing for Functional Analysis: Assessing RNAi as a Tool for Manipulation of Gene Expression Carmit Ziv and Oded Yarden Part II: Detection and Quantification of Fungi 7. Analysis of Fungal Gene Expression by Real Time Quantitative PCR Shahar Ish-Shalom and Amnon Lichter 8. Identification of Differentially Expressed Fungal Genes in planta by Suppression Subtraction Hybridization (SSH) Benjamin A. Horwitz and Sophie Lev 9. Quantification of Fungal Infection of Leaves with Digital Images and Scion Image Software Paul H. Goodwin and Tom Hsiang 10. Expression Profiling of Fungal Genes During Arbuscular Mycorrhiza Symbiosis Establishment Using Direct Fluorescent In situ RT-PCR Pascale M. A. Seddas-Dozolme, Christine Arnould, Marie Tollot, Elena Kuznetsova, and Vivienne Gianinazzi-Pearson 11. Application of Laser Microdissection to Study Plant-Fungal Pathogen Interactions John Fosu-Nyarko, Michael G.K. Jones, and Zhaohui Wang 12. Multiplex Gene Expression Analysis by TRAC in Fungal Cultures Jari J. Rautio 13. Amplification of Fungal Genomes Using Multiple Displacement Amplification Simon J. Foster and Brendon J. Monahan Part III: Microscopy and Protein Analysis 14. Biochemical Methods Used to Study the Gene Expression and Protein Complexes in the Filamentous Fungus Neurospora crassa Jinhu Guo, Guocun Huang, Joonseok Cha, and Yi Liu 15. Measuring Protein Kinase and Sugar Kinase Activity in Plant Pathogenic Fusarium Species Burton H. Bluhm and Xinhua Zhao 16. A Detailed Protocol for Chromatin Immunoprecipitation (ChIP) in the Yeast Saccharomyces cerevisiae Melanie Grably and David Engelberg 17. A Method to Visualize the Actin and Microtubule Cytoskeleton by Indirect Immunofluorescence Flora Banuett 18. Fluorescence in situ Hybridization for Molecular Cytogenetic Analysis in Filamentous Fungi Dai Tsuchiya and Masatoki Taga 19. Live-Cell Imaging of Microtubule Dynamics in Hyphae of Neurospora crassa Maho Uchida, Rosa R. Mourino-Perez, and Robert W. Roberson 20. Methods to Detect Apoptotic-Like Cell Death in Filamentous Fungi Camile P. Semighini and Steven D. Harris 21. Evaluation of Antifungal Susceptibility Using Flow Cytometry Cidalia Pina-Vaz and Acacio Goncalves Rodrigues 22. Preparation of Fungi for Ultrastuctural Investigations and Immunogoldlabelling Gerd Hause and Simone Jahn 23. Split-EGFP Screens for the Detection and Localization of Protein-Protein Interactions in Living Yeast Cells Emma Barnard and David J. Timson