Plant Argonaute Proteins

Methods and Protocols

Buch | Hardcover

297 Seiten

2017 | 1st ed. 2017
Humana Press Inc. (Verlag)
9781493971640 (ISBN)

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This volume provides the reader with step–by–step methods to study plant ARGONAUTE (AGO) functions. Chapters in Plant Argonaute Poteins: Methods and Protocols summarize the main biochemical, molecular and cytological methods to study AGO–small RNA complexes and their interactions with target RNAs, AGO subcellular localization, AGO association with polysomes, AGO role in meiosis and DNA repair, methods for the identification, cloning and characterization of AGO genes in different plant species, and non–radioactive protocols for small RNA detection. Finally, a series of chapters describing computational methods to study plant AGO function and evolution are provided. 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 troubleshootingand avoiding known pitfalls.

Authoritative and practical, Plant Argonaute Proteins: Methods and Protocols aims to deliver the most complete and updated list of protocols to study plant AGO function.

Plant ARGONAUTEs: Features, Functions and Unknowns.- Analysis of the Uridylation of Both ARGONAUTE–Bound MiRNAs and 5’ Cleavage Products of their Target RNAs in Plants.- In Vitro Formation of Plant RNA-Induced Silencing Complexes Using an Extract of Evacuolated Tobacco Protoplasts.- In vitro Analysis of ARGONAUTE–Mediated Target Cleavage and Translational Repression in Plants.- Establishment of an In Vivo ARGONAUTE Reporter System in Plants.- Immunoprecipitation and High–throughput Sequencing of ARGONAUTE–Bound RNAs from Plants.- Identification of ARGONAUTE/Small RNA Cleavage Sites by Degradome Sequencing.- Dissecting the Sub–Nuclear Localization Patterns of ARGONAUTE Proteins and Other Components of the RNA–Directed DNA Methylation Pathway in Plants.- Detection of ARGONAUTE 1 Association with Polysomes in Arabidopsis thaliana.- Functional Analysis of Arabidopsis ARGONAUTEs in Meiosis and DNA Repair.- Isolation and Characterization of ARGONAUTE Mutants in Chlamydomonas.- ARGONAUTE genes in Salvia miltiorrhiza: Identification, Characterization and Genetic Transformation.- Cloning and Characterization of ARGONAUTE Genes in Tomato.- Non–Radioactive Detection of Small RNAs Using Digoxygenin–Labeled Probes.- Non–Radioactive Plant Small RNA Detection Using Biotin–Labeled Probes.- Computational Analysis of Genome–Wide ARGONAUTE–Dependent DNA.- Structural and Functional Characterization of Plant ARGONAUTE MID Domains.- Identification and Analysis of WG/GW ARGONAUTE–Binding Domains.- Identification and Evolutionary Characterization of ARGONAUTE-Binding Platforms.- Phylogenetic and Evolutionary Analysis of Plant ARGONAUTES.

Erscheinungsdatum	06.07.2017
Reihe/Serie	Methods in Molecular Biology ; 1640
Zusatzinfo	27 Illustrations, color; 20 Illustrations, black and white; XI, 297 p. 47 illus., 27 illus. in color.
Verlagsort	Totowa, NJ
Sprache	englisch
Maße	178 x 254 mm
Themenwelt	Naturwissenschaften ► Biologie ► Botanik
Schlagworte	Affinity purification • ARGONAUTEs • Mass Spectrometry • RIP-seq • target RNA
ISBN-13	9781493971640 / 9781493971640
Zustand	Neuware