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TitleRiboswitches [electronic resource] : Methods and Protocols
Author edited by Alexander Serganov
ImprintTotowa, NJ : Humana Press : Imprint: Humana Press, 2009
Connect tohttp://dx.doi.org/10.1007/978-1-59745-558-9
Descript online resource

SUMMARY

The revolutionary discoveries of RNA interference and bacterial regulatory RNAs led to further breakthroughs such as the identification of riboswitches and related RNA sensors, mRNA regions capable of alternating their conformations in response to the presence of cellular metabolites and other physical or chemical cues. In Riboswitches: Methods and Protocols, expert researchers provide comprehensive and up-to-date coverage of various methods used to study riboswitches and other RNAs involved in gene expression control. Examining biochemical and modern biophysical techniques, the volume focuses on mRNAs responding to small organic molecules but expands the definition of a riboswitch to incorporate classes of RNA that undergo conformational transitions in response to other stimuli in order to control the expression of genes. 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Cutting-edge and authoritative, Riboswitches: Methods and Protocols promises to inspire both novices and experts working in a wide range of contemporary biological areas who wish to further develop their RNA methodology


CONTENT

Predicting Riboswitch Regulation on a Genomic Scale -- Enzymatic Ligation Strategies for the Preparation of Purine Riboswitches with Site-Specific Chemical Modifications -- Application of Fluorescent Measurements for Characterization of Riboswitch–Ligand Interactions -- Transcriptional Approaches to Riboswitch Studies -- Kinetics of Riboswitch Regulation Studied By In Vitro Transcription -- Molecular Basis of RNA-Mediated Gene Regulation on the Adenine Riboswitch by Single-Molecule Approaches -- Methods for Analysis of Ligand-Induced RNA Conformational Changes -- Monitoring RNA–Ligand Interactions Using Isothermal Titration Calorimetry -- Preparation and Crystallization of Riboswitch–Ligand Complexes -- Crystallization of the glmS Ribozyme-Riboswitch -- Riboswitch Conformations Revealed by Small-Angle X-Ray Scattering -- Time-Resolved NMR Spectroscopy: Ligand-Induced Refolding of Riboswitches -- Analysis of the RNA Backbone: Structural Analysis of Riboswitches by In-Line Probing and Selective 2’-Hydroxyl Acylation and Primer Extension -- Identification of Metabolite–Riboswitch Interactions Using Nucleotide Analog Interference Mapping and Suppression -- RNA-Dependent RNA Switches in Bacteria -- Probing mRNA Structure and sRNA–mRNA Interactions in Bacteria Using Enzymes and Lead(II) -- Structural Probing of RNA Thermosensors -- Ribosomal Initiation Complexes Probed by Toeprinting and Effect of trans-Acting Translational Regulators in Bacteria -- Isolation and Characterization of the Heat Shock RNA 1 -- Analysis of tRNA-Directed Transcription Antitermination in the T Box System In Vivo -- In Vitro Selection of Conformational Probes for Riboswitches -- A Green Fluorescent Protein (GFP)-Based Plasmid System to Study Post-Transcriptional Control of Gene Expression In Vivo -- High-Throughput Screens to Discover Synthetic Riboswitches -- A Mammalian Cell-Based Assay for Screening Inhibitors of RNA Cleavage -- In Vitro Selection of glmS Ribozymes


Life sciences Gene expression Biochemistry Nucleic acids Life Sciences Biochemistry general Nucleic Acid Chemistry Gene Expression



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