Plant Argonaute Proteins

This volume provides the reader with stepbystep 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 AGOsmall 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 nonradioactive 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.

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

Inhalt
Plant ARGONAUTEs: Features, Functions and Unknowns.- Analysis of the Uridylation of Both ARGONAUTEBound 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 ARGONAUTEMediated Target Cleavage and Translational Repression in Plants.- Establishment of an In Vivo ARGONAUTE Reporter System in Plants.- Immunoprecipitation and Highthroughput Sequencing of ARGONAUTEBound RNAs from Plants.- Identification of ARGONAUTE/Small RNA Cleavage Sites by Degradome Sequencing.- Dissecting the SubNuclear Localization Patterns of ARGONAUTE Proteins and Other Components of the RNADirected 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.- NonRadioactive Detection of Small RNAs Using DigoxygeninLabeled Probes.- NonRadioactive Plant Small RNA Detection Using BiotinLabeled Probes.- Computational Analysis of GenomeWide ARGONAUTEDependent DNA.- Structural and Functional Characterization of Plant ARGONAUTE MID Domains.- Identification and Analysis of WG/GW ARGONAUTEBinding Domains.- Identification and Evolutionary Characterization of ARGONAUTE-Binding Platforms.- Phylogenetic and Evolutionary Analysis of Plant ARGONAUTES. <p