Cell-Free Gene Expression

Cell-Free Gene Expression

Author: Ashty S. Karim

Publisher: Humana

Published: 2022-01-06

Total Pages: 435

ISBN-13: 9781071619971

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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 ideal guide for researchers seeking technical methods to current aspects of CFE and related applications.


Cell-Free Protein Expression

Cell-Free Protein Expression

Author: James R. Swartz

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 213

ISBN-13: 3642593372

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Cell-free protein synthesis is coming of age! Motivated by an escalating need for efficient protein synthesis and empowered by readily accessible cell-free protein synthesis kits, the technology is expanding both in the range of feasible proteins and in the ways that proteins can be labeled and modified. This volume follows "Cell-Free Translation Systems", edited by Professor Alexander S. Spirin in 2002. Since then, an impressive collection of new work has emerged that demonstrates a substantial expansion of capability. In this volume, we show that proteins now can be efficiently produced using PCR products as DNA templates and that even membrane proteins and proteins with multiple disulfide proteins are obtained at high yields. Many additional advances are also presented. It is an exciting time for protein synthesis technology.


Cell-free Protein Synthesis

Cell-free Protein Synthesis

Author: Alexander S. Spirin

Publisher: John Wiley & Sons

Published: 2007-12-03

Total Pages: 272

ISBN-13: 9783527316496

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With its detailed description of membrane protein expression, high-throughput and genomic-scale expression studies, both on the analytical and the preparative scale, this book covers the latest advances in the field. The step-by-step protocols and practical examples given for each method constitute practical advice for beginners and experts alike.


Cell-Free Synthetic Biology

Cell-Free Synthetic Biology

Author: Yuan Lu

Publisher: Springer

Published: 2019-09-02

Total Pages: 37

ISBN-13: 9789811311703

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This book describes advanced studies in cell-free synthetic biology, an emerging biotechnology that focuses on cell-free protein synthesis and cell-free systems for fundamental and industrial research in areas such as genetic circuit design, small-molecule synthesis, complicated-macromolecule synthesis, unnatural-macromolecule synthesis, high-throughput screening, artificial cells, and biomaterials. Cell-free synthetic biology is now an integral part of developing fields like nanotechnology, materials science, and personalized medicine. The book discusses the main research directions in the development of cell-free systems, as well as a number of applications of cell-free synthetic biology, ranging from structural biology to the human health industry. It is intended for students and researchers in life sciences, synthetic biology, bioengineering, and chemical engineering.


Cell Biology by the Numbers

Cell Biology by the Numbers

Author: Ron Milo

Publisher: Garland Science

Published: 2015-12-07

Total Pages: 400

ISBN-13: 1317230698

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A Top 25 CHOICE 2016 Title, and recipient of the CHOICE Outstanding Academic Title (OAT) Award. How much energy is released in ATP hydrolysis? How many mRNAs are in a cell? How genetically similar are two random people? What is faster, transcription or translation?Cell Biology by the Numbers explores these questions and dozens of others provid


Cell-Free Translation Systems

Cell-Free Translation Systems

Author: A.S. Spirin

Publisher: Springer Science & Business Media

Published: 2012-12-06

Total Pages: 254

ISBN-13: 3642593798

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This is a unique book that describes the most recent achievements in the methodology of protein biosynthesis under cell-free conditions. Various versions of cell-free protein-synthesizing systems and their applications to production of individual proteins on a preparative scale are reviewed. The most recent, advanced methodologies, such as continuous-exchange and continuous-flow cell-free systems and novel effecting batch-format cell-free procedures, are considered. Special attention is drawn to the possibilities of structural (NMR; X-ray) analysis of various gene expression products with the use of a new generation of cell-free systems.


Recombinant Gene Expression

Recombinant Gene Expression

Author: Paulina Balbas

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 505

ISBN-13: 1592597742

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Since newly created beings are often perceived as either wholly good or bad, the genetic alteration of living cells impacts directly on a symbolic meaning deeply imbedded in every culture. During the earlier years of gene expression research, te- nological applications were confined mainly to academic and industrial laboratories, and were perceived as highly beneficial since molecules that were previously unable to be separated or synthesized became accessible as therapeutic agents. Such were the success stories of hormones, antibodies, and vaccines produced in the bacterium Escherichia coli. Originally this bacterium gained fame among humans for being an unwanted host in the intestine, or worse yet, for being occasionally dangerous and pathogenic. H- ever, it was easily identified in contaminated waters during the 19th century, thus becoming a clear indicator of water pollution by human feces. Tamed, cultivated, and easily maintained in laboratories, its fast growth rate and metabolic capacity to adjust to changing environments fascinated the minds of scientists who studied and modeled such complex phenomena as growth, evolution, genetic exchange, infection, survival, adaptation, and further on—gene expression. Although at the lower end of the complexity scale, this microbe became a very successful model system and a key player in the fantastic revolution kindled by the birth of recombinant DNA technology.


High Throughput Protein Expression and Purification

High Throughput Protein Expression and Purification

Author: Sharon A. Doyle

Publisher: Humana Press

Published: 2010-11-19

Total Pages: 0

ISBN-13: 9781617378218

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Despite exciting advances in genome sequencing, isolating a protein from its expression system in its native form still presents a complex challenge. In High Throughput Protein Expression and Purification: Methods and Protocols, leading scientists detail the most successful protocols currently in use, including various high throughput cloning schemes, protein expression analysis, and production protocols. This volume describes the use of E. coli, insect, and mammalian cells, as well as cell-free systems for the production of a wide variety of proteins, including glycoproteins and membrane proteins, in order to best represent strategies that create and exploit common features to enable simplified cloning, stable expression, and purification of proteins. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters present brief introductions to the subject, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section for tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, High Throughput Protein Expression and Purification: Methods and Protocols is an ideal reference for protein biochemists and all those who wish to apply these easy-to-use protocols to the many applicable fields.


Production of Recombinant Proteins

Production of Recombinant Proteins

Author: Gerd Gellissen

Publisher: John Wiley & Sons

Published: 2006-03-06

Total Pages: 429

ISBN-13: 3527604413

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While the choices of microbial and eukaryotic expression systems for production of recombinant proteins are many, most researchers in academic and industrial settings do not have ready access to pertinent biological and technical information since it is normally scattered throughout the scientific literature. This book closes the gap by providing information on the general biology of the host organism, a description of the expression platform, a methodological section -- with strains, genetic elements, vectors and special methods, where applicable -- as well as examples of proteins produced with the respective platform. The systems thus described are well balanced by the inclusion of three prokaryotes (two Gram-negatives and one Gram-positive), four yeasts, two filamentous fungi and two higher eukaryotic cell systems -- mammalian and plant cells. Throughout, the book provides valuable practical and theoretical information on the criteria and schemes for selecting the appropriate expression platform, the possibility and practicality of a universal expression vector, and on comparative industrial-scale fermentation, with the production of a recombinant Hepatitis B vaccine chosen as an industrial example. With a foreword by Herbert P. Schweizer, Colorado State University, USA: "As a whole, this book is a valuable and overdue resource for a varied audience. It is a practical guide for academic and industrial researchers who are confronted with the design of the most suitable expression platform for their favorite protein for technical or pharmaceutical purposes. In addition, the book is also a valuable study resource for professors and students in the fields of applied biology and biotechnology."