In Vitro Transcription and Translation Protocols

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.