Two Hybrid Technologies

This Methods in Molecular Biology(TM) book offers state-of-the art protocols for the investigation of binary rotein-protein interactions (PPIs), including the classical yeast two-hybrid (Y2H) approach, Y2H variants and other in vivo methods for PPI mapping.

Proteinprotein interactions (PPIs) are strongly predictive of functional relationships among proteins in virtually all processes that take place in the living cell. Therefore, the comprehensive exploration of interactome networks is one of the major goals in systems biology. The aim of Two Hybrid Technologies: Methods and Protocols is to provide a compendium of state-of-the art protocols for the investigation of binary PPIs with the classical yeast two-hybrid (Y2H) approach, Y2H variants and other in vivo methods for PPI mapping. Divided into two convenient sections, the first gives a survey of protocols that are currently employed for Y2H high-throughput screens by different expert labs in the field. Rather than detailing the principles of screening, which have been described previously, the focus is on different implementations of Y2H interactome mapping. The second section of the book considers innovative PPI detection methods that have the potential to emerge as alternative high-throughput methodologies. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Two Hybrid Technologies: Methods and Protocols supplies researchers with a comprehensive toolbox for the identification of biologically relevant protein interactions.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Matrix-Based Yeast Two-Hybrid Screen Strategies and Comparison of Systems.- Array-Based Yeast Two-Hybrid Screens: A Practical Guide.- High Throughput Yeast Two-Hybrid Screening.- A Stringent Yeast Two-Hybrid Matrix Screening Approach for Protein-Protein Interaction Discovery.- High-Throughput Yeast Two-Hybrid Screening of Complex cDNA Libraries.- Virus-Human Cell Interactomes.- Interactome Mapping in Malaria Parasites: Challenges and Opportunities.- Mapping Interactomes with High Coverage and Efficiency Using the Shifted Transversal Design.- Assigning Confidence Scores for Protein-Protein Interactions.- The Intergration and Annotation of the Human Interactome in the UniHI Database.- Gene-Centered Yeast One-Hybrid Assays.- One- plus Two-hybrid System for the Efficient Selection of Missense Mutant Alleles Defective in Protein-Protein Interactions.- Investigation of Membrane Protein Interactions Using the Split-Ubiquitin Membrane Yeast Two-Hybrid System.- Application of the Split-Protein Sensor Trp1 to Protein Interaction Discovery in the Yeast Saccharomyces cerevisiae.- Tetracycline Repressor-Based Mammalian Two-Hybrid Systems.- The Fluorescent Two-Hybrid (F2H) Assay for Direct Analysis of Protein-Protein Interactions in Living Cells.- ArrayMAPPIT: a Screening Platform for Human Protein Interactome Analysis.- MAPPIT as a High-Throughput Screening Assay for Modulators of Protein-Protein Interactions in HIV and HCV.- Integrated Measurement of Split TEV and cis-Regulatory Assays Using EXT Encoded Reporter Libraries.