Fungi are an economic very important class of microbes. Not only do they host a range of versatile enzymes used in industrial applications (biofuels, laundry, food processing), as well do they produce several very important pharmaceutical drugs (statins and penicillins). Moreover, fungal pathogens can cause great damage in agricultural production (Phytophthora and Botrytis) and during mammalian infections (Penicillium marneffei and Candida). Transformation of DNA is used to understand the genetic basis behind these traits. Several different techniques have been developed over the years and readily shown to be decisive methods to improve fungal biotechnology. This book will cover the basics behind the most commonly used transformation methods, as well as associated tools and techniques. Each chapter will provide protocols along with examples to be used in laboratories worldwide.
I. Endogenous DNA: Cell Fusion1. Anastomosis and Heterokaryon FormationMartin Weichert and André Fleißner2. Induction of the Sexual Cycle in Filamentous AscomycetesJos Houbraken and Paul S. Dyer3. What Have We Learned by Doing Transformations in Neurospora tetrasperma ?Durgadas P. KasbekarII. Endogenous DNA: Repetitive Elements4. Repeat-induced Point Mutation: A Fungal-specific, Endogenous Mutagenesis Process James K. Hane, Angela H. Williams, Adam P. Taranto, Peter S. Solomon and Richard P. Oliver5. Calculating RIP Mutation in Fungal Genomes Using RIPCALJames K. Hane6. Fungal Transposable ElementsLinda Paun and Frank Kempken7. In vivo Targeted Mutagenesis in Yeast using TaGTEAMShawn Finney-Manchester and Narendra Maheshri III. Endogenous DNA: RNAi8. RNA Silencing in Filamentous Fungi: from Basic to ApplicationsNguyen Bao Quoc and Hitoshi Nakayashiki9. RNAi-mediated Gene Silencing in the Beta-lactam Producers Fungi Penicillium chrysogenum and A
cremonium chrysogenum Carlos García-Estrada and Ricardo V. Ullán10. Controlling Fungal Gene Expression Using the Docyclyline-Dependent Tet-ON System in Aspergillus fumigatus Michaela Dümig and Sven KrappmannIV. Tools & Applications: Selection Markers and Vectors11. Expanding the Repertoire of Selectable Markers for Aspergillus TransformationKhyati Dave, V. Lakshmi Prabha, Manmeet Ahuja, Kashyap Dave, S. Tejaswini and Narayan S. Punekar12. Arginase ( agaA ) as a Fungal Transformation MarkerKashyap Dave, Manmeet Ahuja, Jayashri, T.N., Rekha Bisht Sirola, Khyati Dave and Narayan S. Punekar13. Transformation of Ascomycetous Fungi using Autonomously Replicating VectorsTakeo Shimizu and Satoko Kanematsu14. A Recyclable and Bidirectionally Selectable Marker System for Transformation of Trichoderma Thiago M. Mello-de-Sousa, Robert L. Mach and Astrid R. Mach-Aigner15. Split Marker-Mediated Transformation and Targeted Gene Disruption in Filamentous FungiKuang-Ren
Chung and Miin-Huey LeeV. Tools & Applications: High Throughput Experimentation16. Integrated Automation for Continuous High-Throughput Synthetic Chromosome Assembly and Transformation to Identify Improved Yeast Strains for Industrial Production of Biofuels and Bio-based ChemicalsStephen R. Hughes and Steven B. Riedmuller17. Imaging Flow Cytometry and High-throughput Microscopy for Automated Macroscopic Morphological Analysis of Filamentous FungiAydin Golabgir, Daniela Ehgartner, Lukas Neutsch, Andreas E. Posch, Peter Sagmeister and Christoph Herwig18. Yeast Cell Electroporation in Droplet-Based Microfluidic ChipQiuxian Cai and Chunxiong Luo19. Identification of T-DNA Integration Sites: TAIL-PCR and Sequence AnalysisJaehyuk Choi, Junhyun Jeon and Yong-Hwan LeeVI. Tools & Applications: Comprehensive Approaches in Selected Fungi20. Genetic and Genomic Manipulation in Aspergillus niger Adrian Tsang, Annie Bellemare, Corinne Darmond and Janny Bakhuis21. Gene
tic Manipulation of Meyerozyma guilliermondii Nicolas Papon,Yuriy R. Boretsky, Vincent Courdavault, Marc Clastre and Andriy A. Sibirny
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