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This book reviews the development of cell-free production platforms and offers an authoritative perspective of the latest advances and methodologies in cell-free production systems. Readers will discover the biomanufacturing potential of in vitro biotransformation (ivBT) employing purified cascade multi-enzymes, the development of hydrogel-based multi-enzymatic systems for biosynthesis, and novel insights into the optimization of biocatalytic processes. Additionally, the book explores the cell-free production and regeneration of cofactors, shedding light on strategies to enhance the efficiency and sustainability of cellular processes.In this book, particular attention is given to the progress of cell-free in vitro evolution techniques for optimizing enzyme performance, and the book also presents the integration of rapid and finely-tuned expression systems for deployable sensing applications, revolutionizing the field of biosensing. The synthesis and electrophysiological analysis of multipass voltage-gated ion channels tethered in microsomal membranes are explored, providing a deep understanding of cellular function at the molecular level. Lastly, the book covers compartmentalized cell-free expression systems for building synthetic cells, showcasing the potential for constructing artificial cellular systems with unique functionalities.Given its breadth, this book appeals to academics, researchers, and professionals interested in the forefront of biotechnology, and together with the companion volume ¿Cell-free Macromolecular Synthesis¿, both books highlight the research progresses on the basic and applied research of cell-free production systems in the last few years, being invaluable resources in the field.Chapter ¿Cell-free synthesis and electrophysiological analysis of multipass voltage-gated ion channels tethered in microsomal membranes¿ is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
This detailed volume explores perspectives and methods using cell-free expression (CFE) to enable next-generation synthetic biology applications. The first section focuses on tools for CFE systems, including a primer on DNA handling and reproducibility, as well as methods for cell extract preparation from diverse organisms and enabling high-throughput cell-free experimentation. The second section provides an array of applications for CFE systems, such as metabolic engineering, membrane-based and encapsulated CFE, cell-free sensing and detection, and educational kits. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Cell¿Free Gene Expression: Methods and Protocols serves as an idealguide for researchers seeking technical methods to current aspects of CFE and related applications.
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