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Full Description
This book presents current molecular biology methods involving Aspergillus fumigatus for researchers in mycology, fungal genetics, and medical microbiology looking to deepen their understanding of this species of fungus at the molecular level. Sections of the book explore approaches to CRISPR-based genome editing, methods for identification, genotyping, and detection, assays for gene regulation, protocols for electrophoretic mobility shift assays and chromatin immunoprecipitation, as well as models to study host-pathogen interactions and methods to investigate the fungal cell wall and secreted components. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Aspergillus fumigatus: Methods and Protocols serves as an ideal guide for scientists attempting to shorten the time spent on routine problems at the bench and move more quickly from idea to results.
Contents
Cas9 Ribonucleoproteins (RNPs) for Gene Deletion.- Generation of Seamless Point Mutations with Cas9 RNP and pTel-hygR Plasmid in Aspergillus fumigatus.- Marker-Free CRISPR/Cas9 RNP Transformation for Gene Disruption in Aspergillus fumigatus.- CRISPR-Cas9 and Microhomology-Mediated End-Joining for Biosynthetic Pathway Reconstruction in Aspergillus fumigatus.- PCR Typing and Genome Sequencing of Aspergillus fumigatus.- Isolation of Nucleic Acids from Aspergillus fumigatus for Sequencing.- Methods for Large-Scale Intraspecific Analyses of Single Nucleotide Polymorphisms in the Mitochondrial Genomes of Aspergillus fumigatus.- Development and Validation of a Real-Time PCR Assay for the Detection of Aspergillus fumigatus on Panther Fusion System.- RAPD-PCR for Typing and Identification of Aspergillus fumigatus.- Electrophoretic Mobility Shift Assay in Aspergillus fumigatus.- Chromatin Immunoprecipitation in Aspergillus fumigatus.- Micropatterning Substrates Using Deep UV Lithography: A Model to Study Host-Pathogen Interactions of A. fumigatus with Adherent Cells.- Bronchoalveolar Lavage Fluid Collection in Invasive Pulmonary Aspergillosis Murine Model for Downstream Proteomic Analysis.- Galleria mellonella to Study Aspergillus Infections and Antifungal Efficacy by Traditional Readouts and Novel BL-Imaging Techniques.- Molecular Biosensing of the Aspergillus fumigatus Cell Wall Integrity Pathway Using the (p)agsA::luc (Luciferase) Reporter System.- Evaluation of Aspergillus fumigatus Metabolic Activity In Vitro: A Comparison Between Resazurin and XTT Methods.- Aspergillus fumigatus Cell Wall: Current Advances in Structural Elucidation Methods.- Isolation and Purification of Extracellular Vesicles from Aspergillus fumigatus.
