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Now a major approach in the identification of certain gene product functions, reverse chemical genetics allows the identification of target genes to precede phenotypic changes created by specific small molecules. In Reverse Chemical Genetics: Methods and Protocols, expert authors introduce fundamental resources, including databases for reverse chemical genetics, as well as the modification of these cDNA resources by the most recent molecular technologies. The book explores newly established methods in addition to potentially applicable techniques that make use of specific small molecules, plus functional assays utilizing these molecules, while focusing on methods that make feasible the functional discovery and comprehensive knowledge of particular target genes. As a volume in the highly successful Methods in Molecular Biology(TM) series, 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 cutting-edge, Reverse Chemical Genetics: Methods and Protocols promises to inspire researchers to conduct their own experiments in order to progress this vital field of study.
List of contents
The Importance of Fully-Characterized Genomic Resources and Informatics for Reverse Chemical Genetics.- ORFeome Cloning.- Systematic Cloning of an ORFeome Using the Gateway System.- High-Throughput Construction of ORF Clones for Production of the Recombinant Proteins.- Computational Overview of GPCR Gene Universe to Support Reverse Chemical Genomics Study.- High-Performance Gene Expression Module Analysis Tool and Its Application to Chemical Toxicity Data.- Human Protein Reference Database and Human Proteinpedia as Discovery Tools for Systems Biology.- Modification of cDNA Resources Utilized for Reverse Chemical Genetics.- High-Throughput Production of the Recombinant Proteins Expressed in Escherichia coli Utilizing cDNA Resources.- An Insect Cell-Free System for Recombinant Protein Expression Using cDNA Resources.- Recombinant Protein Production by a Kaiko-Baculovirus System.- Pulse-Chase Experiment for the Analysis of Protein Stability in Cultured Mammalian Cells by Covalent Fluorescent Labeling of Fusion Proteins.- The Refinement of Established Methods by Recent Innovations.- Application of 2D-DIGE in Cancer Proteomics Toward Personalized Medicine.- Fully Automated Two-Dimensional Electrophoresis System for High-Throughput Protein Analysis.- Large-Scale Whole Mount In Situ Hybridization of Mouse Embryos.- Identification of the Specific Binding Proteins of Bioactive Small Compound Using Affinity Resins.- Potential Applicable Techniques for Reverse Chemical Genetics.- Protein Microarrays: Effective Tools for the Study of Inflammatory Diseases.- Preparation of Highly Sensitive Protein Array Using Reactive Polymer.- High-Throughput SPR Biosensor.- Drug Discovery Through Functional Screening in the Drosophila Heart.- Permeable Cell Assay: A Method for High-ThroughputMeasurement of Cellular ATP Synthetic Activity.- Novel Methodology for Immobilization of Biomolecules on the Surface of a Photoresponsible Polymer Containing Azobenzene Moiety.
Summary
Now a major approach in the identification of certain gene product functions, reverse chemical genetics allows the identification of target genes to precede phenotypic changes created by specific small molecules. In Reverse Chemical Genetics: Methods and Protocols, expert authors introduce fundamental resources, including databases for reverse chemical genetics, as well as the modification of these cDNA resources by the most recent molecular technologies. The book explores newly established methods in addition to potentially applicable techniques that make use of specific small molecules, plus functional assays utilizing these molecules, while focusing on methods that make feasible the functional discovery and comprehensive knowledge of particular target genes. As a volume in the highly successful Methods in Molecular Biology™ series, 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 cutting-edge, Reverse Chemical Genetics: Methods and Protocols promises to inspire researchers to conduct their own experiments in order to progress this vital field of study.
