mRNA Processing and Metabolism

Methods and Protocols

Buch | Softcover

270 Seiten

2010 | Softcover reprint of hardcover 1st ed. 2004
Humana Press Inc. (Verlag)
978-1-61737-434-0 (ISBN)

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Cells possess a wealth of posttranscriptional control mechanisms that impact on every conceivable aspect of the life of an mRNA. These processes are intimately intertwined in an almost baroque manner, where promoter context influences the recruitment of splicing factors, where the majority of pre-mRNAs undergo alternative splicing, and where proteins deposited during nuclear processing impact distal cytoplasmic processing, translation, and decay. If there is a unifying theme to mRNA Processing and Metabolism: Methods and Protocols, it is that mRNA processing and metabolism are integrated processes. Many of the techniques used to study mRNA have been described in a previous volume of this series (RNA–Protein Interaction Protocols, Susan Haynes, ed.) and specialized methods journals. In selecting topics for mRNA Processing and Metabolism: Methods and Protocols, I sought input on new and novel techniques and approaches that build on this foundation using technological advances in microscopy, whole genome sequencing, microarrays, mass spectrometry, fluorescent detection methodologies, and RNA interference. I have tried not to bias this book toward any single model organism, and approaches described in the various chapters use yeast, Drosophila, Xenopus, mice, plants, and cultured mammalian cells.

Using Chromatin Immunoprecipitation to Map Cotranscriptional mRNA Processing in Saccharomyces cerevisiae.- Identifying PhosphoCTD-Associating Proteins.- Imaging Alternative Splicing in Living Cells.- Tobramycin Affinity Tag Purification of Spliceosomes.- Using Single-Strand Conformational Polymorphism Gel Electrophoresis to Analyze Mutually Exclusive Alternative Splicing.- Methods for the Analysis of Adenosine-to-Inosine Editing in RNA.- Assaying Nuclear Messenger RNA Export in Human Cells.- RNA Unwinding Assay for DExD/H-Box RNA Helicases.- Approaches for Monitoring Nuclear Translation.- Immunopurification and Analysis of Protein and RNA Components of mRNP in Mammalian Cells.- Gene Expression Analysis of Messenger RNP Complexes.- Using the ?N Peptide to Tether Proteins to RNAs.- An Efficient System for Cap- and Poly(A)-Dependent Translation In Vitro.- A Poly(A) Tail-Responsive In Vitro System for Cap- or IRES-Driven Translation From HeLa Cells.- Assessing Messenger RNA Decapping in Cellular Extracts.- Analysis of RNA Exonucleolytic Activities in Cellular Extracts.- Application of Ligation-Mediated Reverse Transcription Polymerase Chain Reaction to the Identification of In Vivo Endonuclease-Generated Messenger RNA Decay Intermediates.- Biochemical Dissection of RNA Silencing in Plants.- RNA Interference of mRNA Processing Factors in Drosophila S2 Cells.- RNA Interference by Short Hairpin RNAs Expressed in Vertebrate Cells.

"...the shear diversity of current techniques on offer in this book makes it a valuable resource not only for those studying mRNA processing and metabolism but for those interested in RNA or RNA-protein interactions in general." - ChemBioChem

Erscheint lt. Verlag	10.11.2010
Reihe/Serie	Methods in Molecular Biology ; 257
Zusatzinfo	XVI, 270 p.
Verlagsort	Totowa, NJ
Sprache	englisch
Maße	152 x 229 mm
Themenwelt	Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie
Themenwelt	Naturwissenschaften ► Biologie ► Zellbiologie
ISBN-10	1-61737-434-2 / 1617374342
ISBN-13	978-1-61737-434-0 / 9781617374340
Zustand	Neuware