E Coli Gene Expression Protocols

Author : Peter E. Vaillancourt
Publisher : Springer Science & Business Media
ISBN 13 : 1592593011
Total Pages : 347 pages
Book Rating : 4.19/5 ( download)

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Download or read book E. coli Gene Expression Protocols written by Peter E. Vaillancourt and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 347 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.

Author : Thomas C. Evans, Jr.
Publisher : Humana Press
ISBN 13 : 9781617379680
Total Pages : 310 pages
Book Rating : 4.89/5 ( download)

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Download or read book Heterologous Gene Expression in E.coli written by Thomas C. Evans, Jr. and published by Humana Press. This book was released on 2011-08-24 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein expression in a heterologous host is a cornerstone of biomedical research and of the biotechnology industry. Despite the advanced state of protein expression technology improvements are still needed. For example, membrane proteins constitute a significant percentage of the total cellular proteins but as a class are very difficult to overexpress, especially in a heterologous host. The ideal host would have the ability to express any protein, with relevant post-translational modifications, and be as easy to work with as E. coli. In Heterologous Gene Expression in E. coli: Methods and Protocols, expert scientists intimately familiar with the relevant techniques offer chapters that greatly expand the utility of this expression host. The contributions in this detailed volume describe methods, for example, to successfully express proteins in E. coli that would otherwise form aggregates in this host, to add post-translational modifications, to incorporate non-standard amino acid residues or moieties into E. coli expressed proteins, to identify binding partners, and to express membrane proteins. Written in the highly successful Methods in Molecular BiologyTM format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Heterologous Gene Expression in E. coli: Methods and Protocols seeks to familiarize the researcher with the myriad of E. coli expression strains available and move E. coli closer to that ideal of the perfect host.

Author : John M. Walker
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.96/5 ( download)

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Download or read book Methods in Molecular Biology: E. coli gene expression protocols written by John M. Walker and published by . This book was released on 1984 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Author : Paulina Balbas
Publisher : Springer Science & Business Media
ISBN 13 : 1592597742
Total Pages : 505 pages
Book Rating : 4.41/5 ( download)

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Download or read book Recombinant Gene Expression written by Paulina Balbas and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 505 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Author : Nicola A. Burgess-Brown
Publisher : Humana Press
ISBN 13 : 9781493983285
Total Pages : 429 pages
Book Rating : 4.88/5 ( download)

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Download or read book Heterologous Gene Expression in E.coli written by Nicola A. Burgess-Brown and published by Humana Press. This book was released on 2018-07-20 with total page 429 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E. coli. The topics are largely grouped under four parts: high-throughput cloning, expression screening, and optimization of expression conditions, protein production and solubility enhancement, case studies to produce challenging proteins and specific protein families, as well as applications of E. coli expression. 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, Heterologous Gene Expression in E. coli: Methods and Protocols serves molecular biologists, biochemists and structural biologists, those in the beginning of their research careers to those in their prime, to give both an historical and modern overview of the methods available to express their genes of interest in this exceptional organism.

Author : Peter E. Vaillancourt
Publisher : Humana Press
ISBN 13 : 9781588290083
Total Pages : 347 pages
Book Rating : 4.85/5 ( download)

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Download or read book E. coli Gene Expression Protocols written by Peter E. Vaillancourt and published by Humana Press. This book was released on 2002-10-22 with total page 347 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.

Author : Nicola Casali
Publisher : Springer Science & Business Media
ISBN 13 : 1592594093
Total Pages : 311 pages
Book Rating : 4.92/5 ( download)

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Download or read book E. coli Plasmid Vectors written by Nicola Casali and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 311 pages. Available in PDF, EPUB and Kindle. Book excerpt: A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors, and phagemids. Common downstream applications such as mutagenesis of plasmids, recombinant protein expression, and the use of reporter genes, are also described.

Author : Wayne F. Anderson
Publisher : Springer Nature
ISBN 13 : 1493903543
Total Pages : 344 pages
Book Rating : 4.42/5 ( download)

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Download or read book Structural Genomics and Drug Discovery written by Wayne F. Anderson and published by Springer Nature. This book was released on 2014-07-08 with total page 344 pages. Available in PDF, EPUB and Kindle. Book excerpt: Structural Genomics and Drug Discovery: Methods and Protocols focuses on high throughput structure determination methods and how they can be applied to lay the groundwork for structure aided drug discovery. The methods and protocols that are described can be applied in any laboratory interested in using detailed structural information to advance the initial stages of drug discovery. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Structural Genomics and Drug Discovery: Methods and Protocols seeks to aid scientists in the further study into structural genomics approach as an efficient initial step toward drug discovery and the methods described will be useful to anyone interested in moving in this direction.

Author : Ashty S. Karim
Publisher :
ISBN 13 : 9781071619988
Total Pages : 435 pages
Book Rating : 4.85/5 ( download)

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Download or read book Cell-Free Gene Expression written by Ashty S. Karim and published by . This book was released on 2022 with total page 435 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Author : Eduardo A. Ceccarelli
Publisher : Frontiers E-books
ISBN 13 : 2889192946
Total Pages : 103 pages
Book Rating : 4.46/5 ( download)

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Download or read book Recombinant protein expression in microbial systems written by Eduardo A. Ceccarelli and published by Frontiers E-books. This book was released on 2014-10-02 with total page 103 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the advent of recombinant DNA technology, expressing heterologous proteins in microorganisms rapidly became the method of choice for their production at laboratory and industrial scale. Bacteria, yeasts and other hosts can be grown to high biomass levels efficiently and inexpensively. Obtaining high yields of recombinant proteins from this material was only feasible thanks to constant research on microbial genetics and physiology that led to novel strains, plasmids and cultivation strategies. Despite the spectacular expansion of the field, there is still much room for progress. Improving the levels of expression and the solubility of a recombinant protein can be quite challenging. Accumulation of the product in the cell can lead to stress responses which affect cell growth. Buildup of insoluble and biologically inactive aggregates (inclusion bodies) lowers the yield of production. This is particularly true for obtaining membrane proteins or high-molecular weight and multi-domain proteins. Also, obtaining eukaryotic proteins in a prokaryotic background (for example, plant or animal proteins in bacteria) results in a product that lack post-translational modifications, often required for functionality. Changing to a eukaryotic host (yeasts or filamentous fungi) may not be a proper solution since the pattern of sugar modifications is different than in higher eukaryotes. Still, many advances in the last couple of decades have provided to researchers a wide variety of strategies to maximize the production of their recombinant protein of choice. Everything starts with the careful selection of the host. Be it bacteria or yeast, a broad list of strains is available for overcoming codon use bias, incorrect disulfide bond formation, protein toxicity and lack of post-translational modifications. Also, a huge catalog of plasmids allows choosing for different fusion partners for improving solubility, protein secretion, chaperone co-expression, antibiotic resistance and promoter strength. Next, controlling culture conditions like temperature, inducer and media composition can bolster recombinant protein production. With this Research Topic, we aim to provide an encyclopedic account of the existing approaches to the expression of recombinant proteins in microorganisms, highlight recent discoveries and analyze the future prospects of this exciting and ever-growing field.