An update of the previous edition, In Vitro Transcription and Translation Protocols provides molecular biology laboratories with the most powerful techniques for employing in vitro transcriptions and translation systems. Consisting of sections focusing on technology and applications, topics include continuous-exchange protein synthesizing systems, energy systems for ATP regeneration in cell-free protein synthesis reactions, as well as examining in vitro translation to study HIV protease activity.
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.
...an excellent starting point for those who are starting out to study these important aspects of gene expression and its regulation. - Doody's Health Sciences Book Review Journal The book is of interest not only to experimental endocrinologists and geneticists, but to anyone working on or trying to understand the molecular basis of disease and looking for new paths of healing. - Journal of Pediatric Endocrinology and Metabolism