Main description:

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 (TM) 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.

Practical and hands-on, Fungal Secondary Metabolism: Methods and Protocols encourages new investigators to enter the field and expands upon the expertise and range of skills of those already researching fungal natural products.

Contents:

1. Library Preparation and Data Analysis Packages for Rapid Genome Sequencing

Kyle R. Pomraning, Kristina M. Smith, Erin L. Bredeweg, Lanelle R. Connolly, Pallavi A. Phatale, and Michael Freitag

2. Bioinformatics Approaches and Software for Detection of Secondary Metabolic Gene Clusters

Natalie D. Fedorova, Venkatesh Moktali, and Marnix H. Medema

3. Media and Growth Conditions for Induction of Secondary Metabolite Production

Jens C. Frisvad

4. Manipulating Filamentous Fungus Chemical Phenotypes by Growth on Nutritional Arrays

Jose R. Tormo, Francisco J. Asensio, and Gerald F. Bills

5. The "FERMEX" Method for Metabolite-Enriched Fungal Extracts

Gerald F. Bills, Anne W. Dombrowski, and Michael A. Goetz

6. The Chemical Identification and Analysis of Aspergillus nidulans Secondary Metabolites

James F. Sanchez and Clay C.C. Wang

7. Detection and Purification of Non-Ribosomal Peptide Synthetase Products in Neosartorya fischeri

Beate Wollinsky and Shu-Ming Li

8. Production, Detection, and Purification of Clavine-Type Ergot Alkaloids

Christiane Wallwey and Shu-Ming Li

9. Analysis of Volatile Compounds Emitted by Filamentous Fungi Using Solid Phase Microextraction - Gas Chromatography/Mass Spectrometry (SPME-GC/MS)

Ludmila V. Roze, Randolph M. Beaudry, and John E. Linz

10. Tools for Manipulation of Secondary Metabolism Pathways: Rapid Promoter Replacements and Gene Deletions in Aspergillus nidulans

C. Elizabeth Oakley, Heather Edgerton-Morgan, and Berl R. Oakley

11. Fast and Easy Method for Construction of Plasmid Vectors Using Modified Quick-Change Mutagenesis

Jin Woo Bok and Nancy P. Keller

12. Reconstitution of a Secondary Metabolite Biosynthetic Pathway in a Heterologous Fungal Host

Takayuki Itoh, Tetsuo Kushiro, and Isao Fujii

13. Multi-Copy Genetic Screen in Aspergillus nidulans

Hee Soo Park and Jae-Hyuk Yu

14. Identification of Protein Complexes from Filamentous Fungi with Tandem Affinity Purification

OEzgur Bayram, OEzlem Sarikaya Bayram, Oliver Valerius, Bastian Joehnk, and Gerhard H. Braus

15. Correlating Secondary Metabolite Production with Genetic Changes Using Differential Analysis of 2D NMR Spectra

Ry R. Forseth and Frank C. Schroeder

16. Chromatin Immunoprecipitation (ChIP) Analysis in Filamentous Fungi

Yazmid Reyes-Dominguez, Stefan Boedi, and Joseph Strauss

17. Targeted Proteomics for Metabolic Pathway Optimization

Tanveer S. Batth, Jay D. Keasling, and Christopher J. Petzold

18. Investigation of In Vivo Protein Interactions in Aspergillus Spores

Kwang Cheol Jeong and Jae-Hyuk Yu

19. Purification of a Vesicle-Vacuole (V) Fraction from Aspergillus

Anindya Chanda, Ludmila V. Roze, and John E. Linz

20. Use of the Hollow Fiber Assay for the Discovery of Novel Anticancer Agents from Fungi

Cedric J. Pearce, Daniel D. Lantvit, Qi Shen, David Jarjoura, Xiaoli Zhang, Nicholas H. Oberlies, David J. Kroll, Mansukh C. Wani, Jimmy Orjala, Djaja D. Soejarto, Norman R. Farnsworth, Esperanza J. Carcache de Blanco, James R. Fuchs, A. Douglas Kinghorn, and Steven M. Swanson

21. Molecular Analysis of Fungal Gene Expression upon Interkingdom Competition with Insects

Ulrike Fohgrub and Frank Kempken

Jose R. Tormo, Francisco J. Asensio, and Gerald F. Bills