About the Book
Highly skilled experimentalists provide a comprehensive series of commonly used, as well as specialized, techniques for analyzing how proteins and RNA interact. Richly detailed and readily reproducible, these methods enable researchers to analyze the structural details of an RNA protein interaction, to determine in detail what parts of the protein and RNA are in close contact, and to isolate RNP complexes from cells. There are also in vitro and in vivo methods and protocols for assaying the effects of proteins and RNP complexes on mRNA metabolism. The methods assume only a knowledge of basic molecular biology, biochemistry, and cell culture, and can be readily adapted to other systems. RNA Protein Interaction Protocols offers a wide-ranging collection of up-to-date experimental methods that will assure productive and reproducible results for all investigators in the field.
Table of Contents:
Labeling and Purification of RNA Synthesized by In Vitro Transcription, Paul A. Clarke. Joining RNA Molecules with T4 DNA Ligase, Melissa J. Moore. RNA-Protein Crosslinking with Photoreactive Nucleotide Analogs, Michelle M. Hanna, Lori Bentsen, Michael Lucido, and Archana Sapre. The Methylene Blue Mediated Photocrosslinking Method for Dectection of Proteins that Interact with Double-Stranded RNA. Zhi-Ren Liu and Christopher W. J. Smith. Probing RNA-Protein Interactions by Psoralen Photocrosslinking, Zhuying Wang and Tariq M. Rana. Identification and Sequence Analysis of RNA-Protein Contact Sites by N-Terminal Sequencing and MALDI-MS, Bernd Thiede, Henning Urlaub, Helga Neubauer, and Brigitte Wittmann-Liebold. RNA Footprinting and Modification Interference Analysis, Paul A. Clarke. Oligonucleotide-Targeted RNase H Protection Analysis of RNA-Protein Complexes, Arthur Gunzl and Albrecht Bindereif. Nitrocellulose Filter Binding for Determination of Dissociation Constants, Kathleen B. Hall and James K. Kranz. Measuring Equilibrium and Kinetic Constants Using Gel Retardation Assays, David R. Setzer. PACE Analysis of RNA-Peptide Interactions, Christopher D. Cilley and James R. Williamson. Detection of Nucleic Acid Interactions Using Surface Plasmon Resonance, Robert J. Crouch, Makoto Wakasa, and Mitsuru Haruki. An Escherichia coli-Based Genetic Strategy for Characterizing RNA Binding Proteins, Chaitanya Jain. Screening RNA-Binding Libraries Using a Bacterial Transcription Antitermination Assay, Kazuo Harada and Alan D. Frankel. In Vitro Genetic Analysis of RNA-Binding Proteins Using Phage Display, Ite A. Laird-Offringa. In Vitro Selection of Aptamers from RNA Libraries, Daniel J. Kenan and Jack D. Keene. Identification of Specific Protein-RNA Target Sites Using Libraries of Natural Sequences, Lucy G. Andrews and Jack D. Keene. Northwestern Screening of Expression Libraries, Paramjeet S. Bagga and Jeffrey Wilusz. Screening Expression Libraries with Solution-Based Assay, Philippa J. Webster and Paul M. Macdonald. An Immunoprecipitation-RNA: rPCR Method for the In Vivo Isolation of Ribonucleoprotein Complexes, Edward Chu, John C. Schmitz, Jingfang Ju, and Sitki M. Copur. Purification and Depletion of RNP Particles by Antisense Affinity Chromatography, Benjamin J. Blencowe and Angus I. Lamond. Purification of U Small Nuclear Ribonucleoprotein Particles, Berthold Kastner and Reinhard Luhrmann. Preparation of Heterogeneous Nuclear Ribonucleoprotein Complexes, Maurice S. Swanson and Gideon Dreyfuss. Preparation of Hela Cell Nuclear and Cytosolic S100 Extracts for In Vitro Splicing, Akila Mayeda and Adrian R. Krainer. Mammalian In Vitro Splicing Assays, Akila Mayeda and Adrian R. Krainer. Yeast Pre-mRNA Splicing Extracts, Stephanie W. Ruby. Prespliceosome and Spliceosome Isolation and Analysis, Laura A. Lindsey and Mariano A. Garcia-Blanco. A Yeast Spliceosome Assay, Stephanie W. Ruby. Defining Pre-mRNA cis Elements that Regulate Cell-Specific Splicing, Thomas A. Cooper. In Vivo SELEX in Vertebrate Cells, Thomas A. Cooper. Purification of SR Protein Splicing Factors, Alan M. Zahler. Processing mRNA 3' Ends In Vitro, Michael J. Imperiale. Analysis of Poly(A) Tail Lengths by PCR: The PAT Assay, Fernando J. Salles and Sidney Strickland. In Vitro Translation Extracts from Tissue Culture Cells, Kazuko Shiroki and Akio Nomoto. Messenger RNA Turnover in Cell-Free Extracts from Higher Eukaryotes, Jeff Ross
Review :
This manual is recommended to investigators with experience in molecular biological research, wishing to become introduced into the field of RNA-protein interactions. This book will also be a valuable source of reference for scientists needing general information about the theoretical basis and current methodological approaches to investigating ribonucleoprotein complexes. - International Journal of Medical Microbiology The coverage of the book is broad and reasonably comprehensive, with chapters arranged in a coherent sequence. Many of the authors are well-known, leading researchers in their field, and describe key contemporary techniques. In these respects the coverage is authoritative and likely to remain relevant for some years, before being overtaken by new technologies and methods represents value for money. - Cell Biology International This is a book which any scientist involved in any kind of cellular and molecular biology will find profitable for his experimental investigations since RNA and proteins are unavoidable matters to solve any problem in Life Sciences. Highly skilled experimentalists provide a comprehensive series of commonly used, as well as specialized techniques for analyzing how proteins and RNA interact. Richly detailed and readily reproducible, these methods enable researchers to analyze the structural details of an RNA-protein interaction, to determine in detail what parts of the protein and RNA are in close contact, and to isolate RNP complexes from cells. - Cellular and Molecular Biology
