This volume, part of the Advances in Molecular Biology series, presents work by pioneers in the field and is the first publication devoted solely to the yeast two-hybrid system. It includes detailed protocols, practical advice on troubleshooting, and suggestions for future development. In addition, it illustrates how to construct an activation domain hybrid library, how to identify mutations that disrupt an interaction, and how to use the system in mammalian cells. Many of the contributors have developed new applications and variations of the technique.

Paul N. MacDonald has assembled a collection of powerful molecular tools for examining and characterizing protein-protein, protein-DNA, and protein-RNA interactions. The techniques range from the most basic (introducing plasmids into yeasts, interaction assays, and recovering the plasmids from yeast), to the most advanced alternative strategies (involving one-hybrid, split two-hybrid, three-hybrid, membrane recruitment systems, and mammalian systems). Methods are also provided for dealing with the well-known problem of artifacts and false positives and for identifying the interacting partners in important biological systems, including the SMAD and nuclear receptor pathways. To ensure ready reproducibility and robust results, each technique is described in step-by-step detail by researchers who employ it regularly.

Proteins are indispensable players in virtually all biological events. The functions of proteins are coordinated through intricate regulatory networks of transient protein-protein interactions (PPIs). To predict and/or study PPIs, a wide variety of techniques have been developed over the last several decades. Many in vitro and in vivo assays have been implemented to explore the mechanism of these ubiquitous interactions. However, despite significant advances in these experimental approaches, many limitations exist such as false-positives/false-negatives, difficulty in obtaining crystal structures of proteins, challenges in the detection of transient PPI, among others. To overcome these limitations, many computational approaches have been developed which are becoming increasingly widely used to facilitate the investigation of PPIs. This book has gathered an ensemble of experts in the field, in 22 chapters, which have been broadly categorized into Computational Approaches, Experimental Approaches, and Others.

This is an in-depth examination of circadian biology, presented by leading researchers in the field. Methods for analysis of rhythmic readouts in select model organisms are included. This cutting-edge collection of protocols is adaptable for research at every level, and represents the huge strides that chronobiologists have made over the past two decades. Circadian biologists at all research levels will realize tremendous benefit from this extraordinary collection.

This volume details protocols that cover the broad arsenal of techniques used to study a secretion system from A to Z. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as reporters to follow effector transport. 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. Authoritative and cutting-edge, Bacterial Protein Secretion Systems: Methods and Protocol aims to provide techniques that are not restricted to the study of secretion systems but are also of specific interest for any researcher interested on multi-protein complexes of the bacterial cell envelope.

Protein microarrays soon will become one of the major tools for realizing the goals of drug discovery research in the post-genome era. The book is aimed at life science researchers and scientists working in drug and pharma industry.

This detailed volume explores various aspects of the dengue virus and its four serotypes (DENV1-4). Beginning with a section on protocols to produce DENV and its proteins that are fundamental for many biophysical, biochemical, immunological, and cellular studies, the book continues by covering protocols to study the interactions between DENV and cellular proteins, DENV immunopathogenesis and diagnosis, as well as recent advances in animal models that can be used in studies of DENV immunopathogenesis and vaccine development. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Dengue Virus: Methods and Protocols serves as an ideal guide for researchers attempting to further understand this critical threat to worldwide public health.

In this first integrated view, practically each of the world's leading experts has contributed to this one and only authoritative resource on the topic. Bringing systems biology to cellular energetics, they address in detail such novel concepts as metabolite channeling and medical aspects of metabolic syndrome and cancer.

During recent years, research has greatly expanded our understanding of the sophisticated molecular network of responses which enable plants to develop, survive and propagate under a wide range of conditions. In Plant Signal Transduction: Methods and Protocols, an international panel of experts provide well-established methods vital to analyzing plant signal transduction on the molecular level. Featuring experimental procedures on several of the most popular model organisms, the volume focuses on in planta analyses and the proteins involved in signal transduction in order to aid with the establishment of laboratory techniques or the modification of the protocols for other plants. As part of the highly successful Methods in Molecular BiologyTM series, the chapters include brief introductions to the subject, lists of necessary materials, readily reproducible laboratory protocols, and tips on trouble-shooting and avoiding known pitfalls. Comprehensive and cutting-edge, Plant Signal Transduction: Methods and Protocols will benefit plant scientists wishing to improve their experimental approaches and delve further into this exciting and important field of study

Over the past century, studies of the budding yeast Saccharomyces cerevisiae have helped to unravel principles of nearly every aspect of eukaryotic cell biologyfrom metabolism and molecular genetics to cell division and differentiation. Thanks to its short generation time, ease of genetic manipulation, and suitability for high-throughput studies, yeast remains the focus of research in a vast number of laboratories worldwide. This laboratory manual provides a comprehensive collection of experimental procedures that continue to make budding yeast an informative model. The contributors describe methods for culturing and genetically modifying yeast, strategies and tools (e.g., gene deletion collections) for functional analyses, approaches for characterizing cell structure and morphology, and techniques to probe the modifications and interactions of various cellular constituents (e.g., using one- and two-hybrid screens). Strategies for studying metabolomics, complex traits, and evolution in yeast are also covered, as are methods to isolate and investigate new strains of yeast from the wild. Several additional chapters are devoted to bioinformatics tools and resources for yeast biology (e.g., the Saccharomyces Genome Database). This manual is therefore an essential resource for all researchers, from graduate level upward, who use budding yeast to explore the intricate workings of cells.