In vitro transcription and translation protocols

Bibliographic Information

In vitro transcription and translation protocols

edited by Martin J. Tymms

(Methods in molecular biology / John M. Walker, series editor, 37)

Humana Press, c1995

Available at  / 13 libraries

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Includes bibliographical references and index

Description and Table of Contents

Description

This book provides molecular biology laboratories with the most powerful techniques for employing in vitro transcription and translation systems. Detailed experimental protocols are provided for prokaryotic transcription and translation systems, along with protocols for the many techniques used in the analysis of eukaryotic transcription. The collection is unique in that it also contains protocols for core techniques that use the products of in vitro transcription and translation systems, such as protein import into mitochondria and chloroplasts and structure-function studies. Also covered are core techniques in the analysis of promoters and transcription factors.

Table of Contents

Transcription In Vitro Using Bacteriophage RNA Polymerases. Subtraction Hybridization cDNA Libraries. Quantitative Measurement of mRNA Using the RNase Protection Assay. Assembly and Transcription of Chromatin Templates Using RNA Polymerase III. Antisense Affinity Depletion of RNP Particles: Application to Spliceosomal snRNPs. In Vitro mRNA Editing Using S100 Extracts. An In Vitro Transcription Assay for Probing Drug-DNA Interactions During Active Transcription of DNA. In Vitro Reconstitution of Progesterone-Dependent RNA Transcription in Nuclear Extracts of Human Breast Carcinoma Cells. Plastid In Vitro Transcription. Microinjection of In Vitro Transcribed RNA and Antisense Oligonucleotides in Mouse Oocytes and Early Embryos to Study the Gain- and Loss-of-Function of Genes. mRNA Translation in Xenopus Oocytes. The Xenopus Egg Extract Translation System. In Vitro Translation Using Rabbit Reticulocyte Lysate. An In Vitro Transcription System from BmN Cells of the Silkworm Bombyx mori. Translation Using a Wheat-Germ Extract. In Vitro Transcription and Translation in a Cell-Free System from Clostridium tetani. Preparation and Use of E. coli S-30 Extracts. Posttranslational Transport and In Thylakoido Integration of Plastid Protein. Import into Isolated Yeast Mitochondria of Radiolabeled Proteins Synthesized In Vitro. Structure-Function Studies Based on In Vitro Expression. s-Like Plastid Transcription Factors. Preparation of HeLa Nuclear Extracts. Purification of the Human TATA-Binding Protein TBP. DNA-Binding Studies Using In Vitro Synthesized Myb Proteins. DNase I Footprinting Using PCR-Generated End-Labeled DNA Probes. Gene Isolation by Screening lgtll Expression Libraries with DNA Binding-Site Probes. Transcriptional Activation Analysis by the Chloramphenicol Acetyl Transferase (CAT) Enzyme Assay. Index.

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