Fungal Secondary Metabolism, Softcover reprint of the original 1st ed. 2012
Methods and Protocols
Methods in Molecular Biology Series, Vol. 944

Filamentous fungi have long been known for their ability to produce an enormous range of unusual chemical compounds known as secondary metabolites, many of which have potentially useful antibiotic or pharmacological properties. Recent focus on fungal genomics coupled with advances in detection and molecular manipulation techniques has galvanized a revitalization of this field.  Fungal Secondary Metabolism: Methods and Protocols is aimed at providing the key methodologies currently in use and necessary for accessing and exploiting the natural product information provided by the genomes of this large and varied kingdom. Written by active researchers in the field, the chapters deal with all the steps necessary, from optimization of fungal culture conditions for metabolite production, through rapid genome sequencing and bioinformatics, and genetic manipulations for functional analysis, to detection and testing of metabolites. In addition, chapters on basic science address approaches to the genetic regulation, protein biochemistry, and cellular localization of the biosynthetic pathways. Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Library Preparation and Data Analysis Packages for Rapid Genome Sequencing.- Bioinformatics Approaches and Software for Detection of Secondary Metabolic Gene Clusters.- Media and Growth Conditions for Induction of Secondary Metabolite Production.- Manipulating Filamentous Fungus Chemical Phenotypes by Growth on Nutritional Arrays.- The “FERMEX” Method for Metabolite-Enriched Fungal Extracts.- The Chemical Identification and Analysis of Aspergillus nidulans Secondary Metabolites.- Detection and Purification of Non-Ribosomal Peptide Synthetase Products in Neosartorya fischeri.- Production, Detection, and Purification of Clavine-Type Ergot Alkaloids.- Analysis of Volatile Compounds Emitted by Filamentous Fungi Using Solid Phase Microextraction - Gas Chromatography/Mass Spectrometry (SPME-GC/MS).- Tools for Manipulation of Secondary Metabolism Pathways: Rapid Promoter Replacements and Gene Deletions in Aspergillus nidulans.- Fast and Easy Method for Construction of Plasmid Vectors Using Modified Quick-Change Mutagenesis.- Reconstitution of a Secondary Metabolite Biosynthetic Pathway in a Heterologous Fungal Host.- Multi-Copy Genetic Screen in Aspergillus nidulans.- Identification of Protein Complexes from Filamentous Fungi with Tandem Affinity Purification.- Correlating Secondary Metabolite Production with Genetic Changes Using Differential Analysis of 2D NMR Spectra.- Chromatin Immunoprecipitation (ChIP) Analysis in Filamentous Fungi.- Targeted Proteomics for Metabolic Pathway Optimization.- Investigation of In Vivo Protein Interactions in Aspergillus Spores.- Purification of a Vesicle-Vacuole (V) Fraction from Aspergillus.- Use of the Hollow Fiber Assay for the Discovery of Novel Anticancer Agents from Fungi.- Molecular Analysis of Fungal Gene Expression upon Interkingdom Competition with Insects.