Cloning, Gene Expression, and Protein Purification

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Author :
Publisher : Oxford University Press, USA
ISBN 13 : 9780195132946
Total Pages : 435 pages
Book Rating : 4.1/5 (329 download)

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Book Synopsis Cloning, Gene Expression, and Protein Purification by : Jennifer Edwards

Download or read book Cloning, Gene Expression, and Protein Purification written by Jennifer Edwards and published by Oxford University Press, USA. This book was released on 2001 with total page 435 pages. Available in PDF, EPUB and Kindle. Book excerpt: On the forefront of modern scientific innovation, Cloning, Gene Expression and Protein Purification: Experimental Procedures and Process Rationale effectively doubles as a laboratory manual for students and a reference book for professional researchers. Designed for advanced undergraduate and beginning graduate students in molecular biology, this unique combination lecture/laboratory resource presents detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein's basic physical properties. This manageable volume includes both theoretical background and practical procedures and is structured around twenty experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on unique chemical properties or substrate affinities. The book describes advanced topics such as the use of antibodies and the techniques developed to transform their structures, as well as combinatorial approaches designed to manipulate the structure and functions of proteins and nucleic acids. Supplemental literature provides a variety of theoretical explanations encouraging a more intuitive understanding of the experimental mechanisms and behaviors of the chemical participants, while also giving students the tools needed to become "capable proactive researchers." Features: � Emphasizes electrophoresis, Southern and Western blotting, and combinatorial techniques � Defines clear reaction mechanisms; stipulates the functions of reagents; and helps students think about the precise consequences of solution and procedural manipulations � Discusses fluorophores, and solvent effects on protein structure � Characterizes plasmids, cDNAs, and antibody probes (available from ATCC) in research literature � Includes carefully selected primary source research literature and articles from current vendor literature � Contains a glossary of unfamiliar phrases and jargon; important summary statements and conclusions are italicized � Provides an alphabetized list of common reagents for rapid reference � Offers an extensive index of concepts and terms � Categorizes helpful and distinctive information into five types of supplemental literature: Innovation/ Insight, Theory/Principle, Process Rationale, Vendor Literature, and Alternative Approaches

Textbook on Cloning, Expression and Purification of Recombinant Proteins

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Author :
Publisher : Springer Nature
ISBN 13 : 9811649871
Total Pages : 315 pages
Book Rating : 4.8/5 (116 download)

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Book Synopsis Textbook on Cloning, Expression and Purification of Recombinant Proteins by : Kakoli Bose

Download or read book Textbook on Cloning, Expression and Purification of Recombinant Proteins written by Kakoli Bose and published by Springer Nature. This book was released on 2022-01-25 with total page 315 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Cloning, Gene Expression, And Protein Purification

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Author :
Publisher : Turtleback Books
ISBN 13 : 9781417633326
Total Pages : pages
Book Rating : 4.6/5 (333 download)

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Book Synopsis Cloning, Gene Expression, And Protein Purification by : Jennifer Edwards

Download or read book Cloning, Gene Expression, And Protein Purification written by Jennifer Edwards and published by Turtleback Books. This book was released on 2001-03-01 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Manipulation and Expression of Recombinant DNA

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Publisher : Elsevier
ISBN 13 : 0080456545
Total Pages : 172 pages
Book Rating : 4.0/5 (84 download)

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Book Synopsis Manipulation and Expression of Recombinant DNA by : Sue Carson

Download or read book Manipulation and Expression of Recombinant DNA written by Sue Carson and published by Elsevier. This book was released on 2005-12-15 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Applications of Chimeric Genes and Hybrid Proteins, Part A: Gene Expression and Protein Purification

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Author :
Publisher : Elsevier
ISBN 13 : 0080496814
Total Pages : 651 pages
Book Rating : 4.0/5 (84 download)

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Book Synopsis Applications of Chimeric Genes and Hybrid Proteins, Part A: Gene Expression and Protein Purification by :

Download or read book Applications of Chimeric Genes and Hybrid Proteins, Part A: Gene Expression and Protein Purification written by and published by Elsevier. This book was released on 2000-10-11 with total page 651 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Gene Cloning and Expression Technologies

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Author :
Publisher : Biotechniques Press
ISBN 13 :
Total Pages : 652 pages
Book Rating : 4.3/5 (97 download)

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Book Synopsis Gene Cloning and Expression Technologies by : Michael P. Weiner

Download or read book Gene Cloning and Expression Technologies written by Michael P. Weiner and published by Biotechniques Press. This book was released on 2002 with total page 652 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Recombinant protein expression in microbial systems

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Author :
Publisher : Frontiers E-books
ISBN 13 : 2889192946
Total Pages : 103 pages
Book Rating : 4.8/5 (891 download)

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Book Synopsis Recombinant protein expression in microbial systems by : Eduardo A. Ceccarelli

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.

Cloning and Expression Vectors for Gene Function Analysis

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Author :
Publisher : Eaton Publishing Company/Biotechniques Books
ISBN 13 :
Total Pages : 302 pages
Book Rating : 4.E/5 ( download)

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Book Synopsis Cloning and Expression Vectors for Gene Function Analysis by : Quinn Lu

Download or read book Cloning and Expression Vectors for Gene Function Analysis written by Quinn Lu and published by Eaton Publishing Company/Biotechniques Books. This book was released on 2001 with total page 302 pages. Available in PDF, EPUB and Kindle. Book excerpt:

High Throughput Protein Expression and Purification

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Publisher : Humana Press
ISBN 13 : 9781603279758
Total Pages : 322 pages
Book Rating : 4.2/5 (797 download)

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Book Synopsis High Throughput Protein Expression and Purification by : Sharon A. Doyle

Download or read book High Throughput Protein Expression and Purification written by Sharon A. Doyle and published by Humana Press. This book was released on 2008-11-01 with total page 322 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Gene Expression Systems

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Publisher : Elsevier
ISBN 13 : 0080532357
Total Pages : 493 pages
Book Rating : 4.0/5 (85 download)

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Book Synopsis Gene Expression Systems by : Joseph M. Fernandez

Download or read book Gene Expression Systems written by Joseph M. Fernandez and published by Elsevier. This book was released on 1998-12-21 with total page 493 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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

Recombinant Protein Expression: Eukaryotic hosts

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Author :
Publisher : Academic Press
ISBN 13 : 0323907385
Total Pages : 384 pages
Book Rating : 4.3/5 (239 download)

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Book Synopsis Recombinant Protein Expression: Eukaryotic hosts by :

Download or read book Recombinant Protein Expression: Eukaryotic hosts written by and published by Academic Press. This book was released on 2021-11-04 with total page 384 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant Protein Expression, Part B, Volume 660 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression

E. coli Gene Expression Protocols

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Author :
Publisher : Springer Science & Business Media
ISBN 13 : 1592593011
Total Pages : 347 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis E. coli Gene Expression Protocols by : Peter E. Vaillancourt

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.

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

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Author :
Publisher : Academic Press
ISBN 13 : 0323901476
Total Pages : 462 pages
Book Rating : 4.3/5 (239 download)

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Book Synopsis Recombinant Protein Expression: Prokaryotic hosts and cell-free systems by :

Download or read book Recombinant Protein Expression: Prokaryotic hosts and cell-free systems written by and published by Academic Press. This book was released on 2021-10-29 with total page 462 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recombinant Protein Expression, Part A, Volume 659 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression

Gene Cloning

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Publisher : Garland Science
ISBN 13 : 1135728925
Total Pages : 612 pages
Book Rating : 4.1/5 (357 download)

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Book Synopsis Gene Cloning by : Julia Lodge

Download or read book Gene Cloning written by Julia Lodge and published by Garland Science. This book was released on 2007-01-24 with total page 612 pages. Available in PDF, EPUB and Kindle. Book excerpt: The ability to successfully clone genes underlies the majority of our knowledge in molecular and cellular biology. Gene Cloning introduces the diverse array of techniques available to clone genes and how they can be used effectively both in the research laboratory, to gain knowledge about the gene, and for use in biotechnology, medicine, the pharmaceutical industry, and agriculture. It shows how cloning genes is an integral part of genomics and underlines its relevance in the post-genomic age, as a tool required to test predictions of gene regulation and function made through bioinformatics. Applications of gene cloning in medicine, both for diagnosis and treatment, and in the pharmaceutical industry and agriculture, are also covered in the book. Gene Cloning takes a fresh approach to teaching molecular and cellular biology and will be a valuable resource to both undergraduates and lecturers of biological and biomedical science courses.

Experimental Manipulation of Gene Expression

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Publisher : Academic Press
ISBN 13 : 1483273970
Total Pages : 330 pages
Book Rating : 4.4/5 (832 download)

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Book Synopsis Experimental Manipulation of Gene Expression by : Masayori Inouye

Download or read book Experimental Manipulation of Gene Expression written by Masayori Inouye and published by Academic Press. This book was released on 2014-06-28 with total page 330 pages. Available in PDF, EPUB and Kindle. Book excerpt: Experimental Manipulation of Gene Expression discusses a wide range of host systems in which to clone and express a gene of interest. The aims are for readers to quickly learn the versatility of the systems and obtain an overview of the technology involved in the manipulation of gene expression. Furthermore, it is hoped that the reader will learn enough from the various approaches to be able to develop systems and to arrange for a gene of particular interest to express in a particular system. The book opens with a chapter on the design and construction of a plasmid vector system used to achieve high-level expression of a particular phage regulatory protein normally found in minute amounts in a phage-infected bacterial cell. This is followed by separate chapters on topics such as high-level expression vectors that utilize efficient Escherichia coli lipoprotein promoter as well as various other portions of the lipoprotein gene Ipp; DNA cloning systems for streptomycetes; and the design and application of vectors for high-level, inducible synthesis of the product of a cloned gene in yeast.

Guide to Protein Purification

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Publisher : Academic Press
ISBN 13 : 0080923178
Total Pages : 915 pages
Book Rating : 4.0/5 (89 download)

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Book Synopsis Guide to Protein Purification by : Richard R Burgess

Download or read book Guide to Protein Purification written by Richard R Burgess and published by Academic Press. This book was released on 2009-11-03 with total page 915 pages. Available in PDF, EPUB and Kindle. Book excerpt: Guide to Protein Purification, Second Edition provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition’s publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and biotechnology and sets a standard for best practices in the field. It relates how these traditional and new cutting-edge methods connect to the explosive advancements in the field. This "Guide to" gives imminently practical advice to avoid costly mistakes in choosing a method and brings in perspective from the premier researchers while presents a comprehensive overview of the field today. Gathers top global authors from industry, medicine, and research fields across a wide variety of disciplines, including biochemistry, genetics, oncology, pharmacology, dermatology and immunology Assembles chapters on both common and less common relevant techniques Provides robust methods as well as an analysis of the advancements in the field that, for an individual investigator, can be a demanding and time-consuming process

Recombinant Gene Expression

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Author :
Publisher : Springer Science & Business Media
ISBN 13 : 1592597742
Total Pages : 505 pages
Book Rating : 4.5/5 (925 download)

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Book Synopsis Recombinant Gene Expression by : Paulina Balbas

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.