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 -- 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 -- Transcription System Using a HeLa Cell Mitochondrial Lysate -- 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 -- ?-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 ?gt11 Expression Libraries with DNA Binding-Site Probes -- Transcriptional activation analysis by the Chloramphenicol Acetyl Transferase (CAT) enzyme assay.

Most laboratories conducting studies that use molecular biology techniques employ in vitro transcription and translation systems as a routine part of their day-to-day research. The commercial availability of purified bacterial RNA polymerase and the availability of robust tra- lation systems has made in vitro systems attractive not only as an alt- native to the in vivo expression of genes, but also as good model systems for studying specific aspects of transcription and translation. Although fairly efficient eukaryotic translation systems have been established for a number of years, reconstitution of transcription in vitro has proved to be more difficult. Recent improvements in fractionation techniques and the cloning of proteins involved in transcription have made this a fast moving area of research. Considerable progress has also been made in recent years in developing in vitro systems to study transcription and translation in chloroplasts and mitochondria, together with systems for the study of protein import. In Vitro Transcription and Translation Protocols provides many detailed experimental procedures for prokaryotic transcription and translation systems, together with protocols for many key techniques used in the analysis of eukaryotic transcription. In keeping with the successful format of preceding volumes of the Methods in Molecular Biology series, step-by-step instructions are provided, together with extensive notes that cover troubleshooting and special tips considered important.