Lin RNA-Protein Interaction Protocols

2. Auflage 2008
ISBN: 978-1-60327-475-3
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 488, 416 Seiten, eBook

Reihe: Methods in Molecular Biology

Due to the vital biological importance of RNA and proteins functioning together within a cell, a protocol volume describing experimental procedures to study their interactions should find a home in many laboratories. RNA-Protein Interaction Protocols, Second Edition updates, complements, and expands upon the popular first edition by providing a collection of cutting-edge techniques developed or refined in the past few years along with tried-and-true methods. The expert contributors explore the isolation and characterization of RNA-protein complexes, the analysis and measurement of RNA-protein interaction, and related novel techniques and strategies. Written in the highly successful Methods in Molecular Biology™ series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, RNA-Protein Interaction Protocols, Second Edition is an ideal guide for researchers continuing the study of this all-important biological partnership.

Zielgruppe

Research

Autoren/Hrsg.

Lin, Ren-Jang

Inhaltsverzeichnis

Isolation of a Sequence-Specific RNA Binding Protein, Polypyrimidine Tract Binding Protein, Using RNA Affinity Chromatography.- An Affinity Oligonucleotide Displacement Strategy to Purify Ribonucleoprotein Complexes Applied to Human Telomerase.- RNA Affinity Tags for the Rapid Purification and Investigation of RNAs and RNA–Protein Complexes.- Assembly and Glycerol Gradient Isolation of Yeast Spliceosomes Containing Transcribed or Synthetic U6 snRNA.- Purification of Ribonucleoproteins Using Peptide-Elutable Antibodies and Other Affinity Techniques.- CLIP: Crosslinking and ImmunoPrecipitation of In Vivo RNA Targets of RNA-Binding Proteins.- Quantitative Analysis of Protein-RNA Interactions by Gel Mobility Shift.- Monitoring Assembly of Ribonucleoprotein Complexes by Isothermal Titration Calorimetry.- Characterization of RNA—Protein Interactions by Phosphorothioate Footprinting and Its Applications to the Ribosome.- In Vivo Analysis of Ribonucleoprotein Complexes Using Nucleotide Analog Interference Mapping.- T7 RNA Polymerase-Mediated Incorporation of 8-N3AMP Into RNA for Studying Protein-RNA Interactions.- A Simple Crosslinking Method, CLAMP, to Map the Sites of RNA-Contacting Domains Within a Protein.- Proteins Specifically Modified With a Chemical Nuclease as Probes of RNA-Protein Interaction.- RNA-Protein Crosslink Mapping Using TEV Protease.- Structural Analysis of Protein-RNA Interactions With Mass Spectrometry.- Analyzing RNA-Protein Crosslinking Sites in Unlabeled Ribonucleoprotein Complexes by Mass Spectrometry.- In Vitro Selection of Random RNA Fragments to Identify Protein-Binding Sites Within Large RNAs.- Immunoprecipitation Analysis to Study RNA-Protein Interactions in Xenopus Oocytes.- Mapping the Regions of RNase P Catalytic RNA That Are Potentially inClose Contact With Its Protein Cofactor.- Quantification of MicroRNAs, Splicing Isoforms, and Homologous mRNAs With the Invader Assay.- Analysis of RNA Structure and RNA-Protein Interactions in Mammalian Cells by Use of Terminal Transferase-Dependent PCR.- Duplex Unwinding and RNP Remodeling With RNA Helicases.- Preparation of Efficient Splicing Extracts From Whole Cells, Nuclei, and Cytoplasmic Fractions.- Designing and Utilization of siRNAs Targeting RNA Binding Proteins.- The use of Saccharomyces cerevisiae proteomic libraries to identify RNA-modifying proteins.