Recombinant Gene Expression

Author : Paulina Balbas,Argelia Lorence
Publisher : Springer Science & Business Media
Page : 505 pages
File Size : 54,9 Mb
Release : 2008-02-04
Category : Science
ISBN : 9781592597741

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Recombinant Gene Expression by Paulina Balbas,Argelia Lorence Pdf

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 : Paulina Balbas,Argelia Lorence
Publisher : Humana Press
Page : 508 pages
File Size : 43,9 Mb
Release : 2010-10-28
Category : Science
ISBN : 1617374601

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Recombinant Gene Expression by Paulina Balbas,Argelia Lorence Pdf

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 : Eduardo A. Ceccarelli,Germán L. Rosano
Publisher : Frontiers E-books
Page : 103 pages
File Size : 51,5 Mb
Release : 2014-10-02
Category : Biotechnology
ISBN : 9782889192946

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Recombinant protein expression in microbial systems by Eduardo A. Ceccarelli,Germán L. Rosano Pdf

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.

Author : Argelia Lorence
Publisher : Humana Press
Page : 649 pages
File Size : 44,6 Mb
Release : 2016-08-23
Category : Medical
ISBN : 1493962213

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Recombinant Gene Expression by Argelia Lorence Pdf

Studies related to recombinant gene expression have brought new advance such as the emergence of the “omics” technologies. While Escherichia coli, Sacharomyces cerevisiae and insect cells continue to be the dominant production platforms of recombinant proteins. In Recombinant Gene Expression: Review and Protocols, Third Edition, expert researchers in the field detail many of the methods now commonly used to study recombinant gene expression. These include methods and techniques for bacteria, lower eukaryotes, fungi, plants and plant cells, and animals and animal cells. Written in the highly successful Methods in Molecular BiologyTM 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, Recombinant Gene Expression: Review and Protocols, Third Edition seeks to aid scientists in the further study of this crucially important research into recombinant gene expression.

Author : Joseph M. Fernandez,James P. Hoeffler
Publisher : Elsevier
Page : 480 pages
File Size : 55,9 Mb
Release : 1998-12-21
Category : Science
ISBN : 9780080532356

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Gene Expression Systems by Joseph M. Fernandez,James P. Hoeffler Pdf

Gene Expression Systems: Using Nature for the Art of Expression offers detailed information on a wide variety of gene expression systems from an array of organisms. It describes several different types of expression systems including transient, stable, viral, and transgenic systems. Each chapter is written by a leader in the field. The book includes timelines and examples for each expression system, and provides an overview of the future of recombinant protein expression. Provides detailed information on expression systems Covers a variety of promoters and host organisms enabling researchers to tailor protocols to their specific needs Includes timelines and examples Compares pros and cons of each method

Author : Bruce Alberts
Publisher : Unknown
Page : 0 pages
File Size : 50,7 Mb
Release : 2002
Category : Cytology
ISBN : 0815332181

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Molecular Biology of The Cell by Bruce Alberts Pdf

Author : Martin C. Schmidt,Joseph C. Glorioso
Publisher : Academic Press
Page : 450 pages
File Size : 50,9 Mb
Release : 1999-07-15
Category : Science
ISBN : UVA:X004346184

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Expression of Recombinant Genes in Eukaryotic Systems by Martin C. Schmidt,Joseph C. Glorioso Pdf

The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences. Key Features * Analysis of Gene Expression * Gene Expression Systems for Functional Analysis * Yeast Expression Systems * RNA-based Control of Recombinant Gene Expression * Small Molecule Control of Gene Expression * Viral Systems for Recombinant Gene Expression

Author : Alan Smith
Publisher : CRC Press
Page : 432 pages
File Size : 45,6 Mb
Release : 1994-11-23
Category : Science
ISBN : 0824795431

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Gene Expression in Recombinant Microorganisms by Alan Smith Pdf

Describing the scientific and commercial applications of microbial recombinant DNA technology, this outstanding, single-source reference offers state-of-the-art reviews of gene expression in the most important classes of recombinant microorganisms-providing numerous examples of the expression of homologous genes or heterologous gene products. Presents a unique collection of safety and regulatory considerations from around the world and addresses specific measures to be taken for large-scale industrial operations!

Author : Varsha Gupta,Manjistha Sengupta,Jaya Prakash,Baishnab Charan Tripathy
Publisher : Springer
Page : 527 pages
File Size : 41,5 Mb
Release : 2016-10-22
Category : Technology & Engineering
ISBN : 9789811008757

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Basic and Applied Aspects of Biotechnology by Varsha Gupta,Manjistha Sengupta,Jaya Prakash,Baishnab Charan Tripathy Pdf

This book explores the journey of biotechnology, searching for new avenues and noting the impressive accomplishments to date. It has harmonious blend of facts, applications and new ideas. Fast-paced biotechnologies are broadly applied and are being continuously explored in areas like the environmental, industrial, agricultural and medical sciences. The sequencing of the human genome has opened new therapeutic opportunities and enriched the field of medical biotechnology while analysis of biomolecules using proteomics and microarray technologies along with the simultaneous discovery and development of new modes of detection are paving the way for ever-faster and more reliable diagnostic methods. Life-saving bio-pharmaceuticals are being churned out at an amazing rate, and the unraveling of biological processes has facilitated drug designing and discovery processes. Advances in regenerative medical technologies (stem cell therapy, tissue engineering, and gene therapy) look extremely promising, transcending the limitations of all existing fields and opening new dimensions for characterizing and combating diseases.

Author : Kakoli Bose
Publisher : Springer Nature
Page : 315 pages
File Size : 40,8 Mb
Release : 2022-01-25
Category : Medical
ISBN : 9789811649875

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Textbook on Cloning, Expression and Purification of Recombinant Proteins by Kakoli Bose Pdf

This book is immensely useful for graduate students as well as researchers to understand the basics of molecular biology and Recombinant DNA Technology. It provides a comprehensive overview of different approaches for the synthesis of recombinant proteins from E. coli including their cloning, expression and purification. Recent advances in genomics, proteomics, and bioinformatics have facilitated the use of Recombinant DNA Technology for evaluating the biophysical and biochemical properties of various proteins. The book starts with an introductory chapter on gene cloning, protein expression and purification and its implication in current research and commercial applications. Each chapter provides a lucid set of principles, tools and techniques for both students and instructors. The protocols described have been aptly exemplified, and troubleshooting techniques have been included to aid better understanding. Moreover, the set of questions at the end of each chapter have been particularly formulated to help effective learning.

Author : William Reznikoff,Larry Gold
Publisher : Elsevier
Page : 390 pages
File Size : 53,6 Mb
Release : 2014-05-20
Category : Science
ISBN : 9781483100807

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Maximizing Gene Expression by William Reznikoff,Larry Gold Pdf

Maximizing Gene Expression focuses on prokaryotic and eukaryotic gene expression. The book first discusses E. coli promoters. Topics include structure analysis, steps in transcription initiation, structure-function correlation, and regulation of transcription initiation. The text also highlights yeast promoters, including elements that select initiation sites, transcription regulation, regulatory proteins, and upstream promoter elements. The text also describes protein coding genes of higher eukaryotes; instability of messenger RNA in bacteria; and replication control of the ColE1-type plasmids. The text then describes translation initiation, including the translation of prokaryotes and eukaryotes. The book puts emphasis on the selective degradation of abnormal proteins in bacteria. Topics include proteins rapidly hydrolyzed in E. coli; intracellular aggregates of abnormal polypeptides; energy requirement and pathway for proteins; proteolytic enzymes in E. coli; and regulation of ion expression. The text also highlights the detection of proteins produced by recombinant DNA techniques and mechanism and practice. The book is a good source of information for readers wanting to study gene expression.

Author : Rocky S. Tuan
Publisher : Unknown
Page : 521 pages
File Size : 40,6 Mb
Release : 1997
Category : Science
ISBN : 1592595480

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Recombinant Gene Expression Protocols by Rocky S. Tuan Pdf

Expert international research workers offer the most up-to-date and comprehensive collection of techniques available for expressing one or more genes of interest. The authoritative contributors describe in depth the vectors used for expression in bacteria, yeast, viruses, nonmammalian higher eukaryotes, mammals, and plants, as well as in transgenic animals. They also provide invaluable detailed information on how to introduce the recombinant gene construct into the appropriate expression host. When combined with a companion volume by the same editor, Recombinant Proteins: Detection and Isolation Protocols, both volumes guide the research worker through the entire journey of recombinant gene expression.

Author : Peter E. Vaillancourt
Publisher : Springer Science & Business Media
Page : 347 pages
File Size : 50,9 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592593019

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E. coli Gene Expression Protocols by Peter E. Vaillancourt Pdf

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 : David L. Hacker
Publisher : Humana Press
Page : 311 pages
File Size : 49,6 Mb
Release : 2018-09-03
Category : Science
ISBN : 1493987291

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Recombinant Protein Expression in Mammalian Cells by David L. Hacker Pdf

This detailed volume explores advances in vector design, DNA delivery, cell cultivation, host cell engineering, and bioprocess optimization within the study of recombinant protein expression in mammalian cells. The majority of the protocols employ either Chinese hamster ovary cells (CHO) or human embryonic kidney 293 cells (HEK293), the workhorses of the field, as the production host; however, the methods can be adapted to other mammalian hosts under the appropriate cell-specific conditions. 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and convenient, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols aims to aid researchers in building on our knowledge of protein structure and function and to speed the discovery of new therapeutic proteins.

Author : Sue Carson,Dominique Robertson
Publisher : Elsevier
Page : 172 pages
File Size : 44,5 Mb
Release : 2005-12-15
Category : Science
ISBN : 9780080456546

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Manipulation and Expression of Recombinant DNA by Sue Carson,Dominique Robertson Pdf

This manual is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology, or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students gain hands-on experience from start to finish in subcloning a gene into an expression vector, through purification of the recombinant protein.The second edition has been completely re-written, with new laboratory exercises and all new illustrations and text, designed for a typical 15-week semester, rather than a 4-week intensive course. The “project approach to experiments was maintained: students still follow a cloning project through to completion, culminating in the purification of recombinant protein. It takes advantage of the enhanced green fluorescent protein—students can actually visualize positive clones following IPTG induction. *Cover basic concepts and techniques used in molecular biology research labs*Student-tested labs proven successful in a real classroom laboratories*Exercises simulate a cloning project that would be performed in a real research lab*"Project" approach to experiments gives students an overview of the entire process*Prep-list appendix contains necessary recipes and catalog numbers, providing staff with detailed instructions