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Full Description
This volume aims at providing readers with protocols to enhance their research for the next decade of phytochrome research. The chapters in this book cover topics such as tests for light-dependent interaction of phytochrome binding proteins using yeast two hybrid assays, in vitro pull down, and co-immunoprecipitation from stable transgenic lines; investigations into phosphorylation, SUMOylation, and ubiquitination of phytochromes and other proteins; a look at how in vivo spectroscopy can be used to measure Pfr levels of phytochrome in planta; identifying hormonal action and co-action in light-regulated processes; and ways to use the CRISPR/Cas9 system to generate Physcomitrella mutant lines. 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.
Cutting-edge and comprehensive, Phytochromes: Methods and Protocols serves as a guideline to researchers who are new to the field, and a stepping stone for experienced researchers looking to expand their knowledge of this evolving field.
Contents
Investigation of Light-Regulated Protein-Protein Interactions using Yeast Two-Hybrid Assays.- Rapid Examination of Phytochrome-PHYTOCHROME INTERACTING FACTOR (PIF) Interaction by In Vitro Co-Immunoprecipitation Assay.- Characterization of Light-Regulated Protein-Protein Interactions by In Vivo Co-Immunoprecipitation (Co-IP) Assays from Plant.- Detection of Phytochrome Phosphorylation in Plants.- Detection of SUMOylated Phytochromes in Plants.- Approaches to Identify Protein Ubiquitination Sites in Plants.- Expression, Purification, and Spectral Characterization of Phytochromes.- In Vivo Spectroscopy.- Basic Phytochrome B Calculations.- Quantitative Analysis of Photobodies.- Regulation of Alternative Splicing by Phytochrome.- Phytochrome-Regulation of Seed Germination.- Shade Avoidance and Neighbor Detection.- A Simple Method for Quantification of Protochlorophyllide in Etiolated Arabidopsis Seedlings.- MeasuringPhytochrome-Dependant Light-Input to the Plant Circadian Clock.- Characterization of Flowering Time Mutants.- Rapid Detection of Hormonal Involvement in Light Responses.- Phytochrome and Light Signaling in Marchantia.- Analysis of Physcomitrella Phytochrome Mutants via Photo- and Polarotropism.- CRISPR/Cas9-Mediated Knockout of Physcomitrella patens Phytochromes.- Phylogenetic Methods to Study Light Signaling.
