Baculovirus and Insect Cell Expression Protocols PDF Download
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Author: David W. Murhammer
Publisher: Methods in Molecular Biology
ISBN: 9781493979783
Category : Medical
Languages : en
Pages : 522
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Book Description
The third edition of this volume expands upon the previous two editions with new and up-to-date methods and protocols. Chapters include step-by-step procedures involved in quantifying cell growth, baculovirus infection and cell metabolism, methods to isolate new cell lines and develop your own serum-free medium, and routine maintenance and storage of insect cell lines and baculoviruses, small- and large-scale recombinant protein production with the BEVS in both insect and mammalian cell culture and in insect larvae, production and characterization of baculoviruses, green fluorescent protein, tubular reactors and RNAi, and baculovirus/insect cell system to study apoptosis and generating envelop-modified baculovirus for gene delivery into mammalian cells. 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, Baculovirus and Insect Cell Expression Protocols, Third Edition aims to not only aid the user in successfully completing the tasks described, but also stimulate the development of improved techniques and new applications of baculoviruses and insect cell culture.
Author: David W. Murhammer
Publisher: Methods in Molecular Biology
ISBN: 9781493979783
Category : Medical
Languages : en
Pages : 522
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Book Description
The third edition of this volume expands upon the previous two editions with new and up-to-date methods and protocols. Chapters include step-by-step procedures involved in quantifying cell growth, baculovirus infection and cell metabolism, methods to isolate new cell lines and develop your own serum-free medium, and routine maintenance and storage of insect cell lines and baculoviruses, small- and large-scale recombinant protein production with the BEVS in both insect and mammalian cell culture and in insect larvae, production and characterization of baculoviruses, green fluorescent protein, tubular reactors and RNAi, and baculovirus/insect cell system to study apoptosis and generating envelop-modified baculovirus for gene delivery into mammalian cells. 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, Baculovirus and Insect Cell Expression Protocols, Third Edition aims to not only aid the user in successfully completing the tasks described, but also stimulate the development of improved techniques and new applications of baculoviruses and insect cell culture.
Author:
Publisher: Academic Press
ISBN: 0323907385
Category : Science
Languages : en
Pages : 384
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Book Description
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
Author: David R. O'Reilly
Publisher:
ISBN: 9780195091311
Category : Baculoviruses
Languages : en
Pages : 366
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Book Description
Baculoviruses have proven to be the most powerful and versatile eukaryotic expression vectors available. This unique laboratory manual is designed to help both beginning and experienced researchers construct and use baculovirus vector systems. It simplifies selection of the most appropriatebaculovirus vector design for a given problem, then describes each step of the implementation process--from vector construction to large-scale protein production. The book provides an understanding of how the vectors work; a biological overview of cells, viruses, plasmids, and promoters; guidelinesfor choosing optimum vectors; protocols for growing insect cells and recombinant viruses; methods of analyzing protein products and scaling up protein production; techniques for producing proteins in insect larvae; and easy-to-use maps charting available expression vectors. This comprehensiveapproach has many benefits for researchers and students alike. It allows them to understand how and why the vector system works and offers a rapid comparison of options for choosing the right virus, plasmid or promoter for vector design and construction, with a minimum amount of lost time. Themanual is an invaluable resource for every individual engaged in the production of proteins for any purpose.
Author: Fredric P. Manfredsson
Publisher:
ISBN: 9781493990658
Category : Electronic books
Languages : en
Pages : 328
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Book Description
This volume discusses protocols, ranging from vector production to delivery methods, used to execute gene therapy applications. Chapters are divided into four parts, and cover topics such as design, construction, and application of transcription activation-like effectors; multi-modal production of adeno-associated virus; construction of oncolytic herpes simplex virus; AAV-mediated gene delivery to the mouse liver; and intrathecal delivery of gene therapeutics by direct lumbar puncture in mice. 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. Comprehensive and authoritative, Viral Vectors for Gene Therapy: Methods and Protocols is a valuable resource for researchers, clinicians, and students looking to utilize viral vectors in gene therapy experiments.
Author: M. Cristina Vega
Publisher: Springer
ISBN: 3319272160
Category : Science
Languages : en
Pages : 378
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Book Description
This book presents advanced expression technologies for the production of protein complexes. Since complexes lie at the heart of modern biology, the expression, purification, and characterization of large amounts of high-quality protein complexes is crucial for the fields of biomedicine, biotechnology, and structural biology. From co-expression in E. coli, yeast, mammalian and insect cells to complex reconstitution from individual subunits, this book offers useful insights and guidance for successful protein expressionists. Across several sections readers will discover existing opportunities for the production of protein complexes in bacterial systems (including membrane proteins and cell-free co-expression), methylotrophic and non-methylotrophic yeasts, protozoa (Leishmania terantolae and Dictyostelium discoideum), baculovirus-infected insect cells, mammalian cells, plants and algae. Complex reconstitution from individually purified subunits or subcomplexes is discussed as a complementary strategy. A last section introduces briefly some of the biophysical and structural characterization techniques for macromolecular complexes using state-of-the-art solution scattering and nuclear magnetic resonance. This work is a guided tour over some of the most powerful and successful protein expression technologies, with a focus on co-expression and high-throughput applications. It is addressed to everyone interested in the production and characterization of macromolecular complexes, from university students who want an accessible description of the major co-expression systems to researchers in biomedicine and the life sciences seeking for an up-to-date survey of available technologies.
Author: Dermot Walls
Publisher: Humana Press
ISBN: 9781607619147
Category : Science
Languages : en
Pages : 527
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Book Description
A prerequisite for elucidating the structure and function of any protein is the prior purification of that protein. This necessity has led to the development of many purification schemes and chromatographic methods for the isolation of native proteins from complex sources. In Protein Chromatography: Methods and Protocols, leading researchers present clear protocol-style chapters that are suitable for newcomers and experts alike. The book opens with vital topics in protein biochemistry, addressing such areas as protein stability and storage, avoiding proteolysis during chromatography, protein quantitation methods including immuno-qPCR, and the contrasting challenges that microfluidics and scale-up production pose to the investigator, and then it segues into key methods involving the generation and purification of recombinant proteins through recombinant antibody production and the tagging of proteins, amongst other means, as well as many variations on classic techniques such as ion-exchange and immunoaffinity chromatography. Written in the highly successful Methods in Molecular BiologyTM series format, protocols 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. Authoritative and up-to-date, Protein Chromatography: Methods and Protocols will greatly aid scientists in establishing these essential techniques in their own laboratories and furthering our understanding of the many imperative functions of proteins.
Author: Dhiraj Kumar
Publisher: Springer
ISBN: 331961343X
Category : Science
Languages : en
Pages : 376
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Book Description
This book provides an overview on the basics in insect molecular biology and presents the most recent developments in several fields such as insect genomics and proteomics, insect pathology and applications of insect derived compounds in modern research. The book aims to provide a common platform for the molecular entomologist to stimulate further research in insect molecular biology and biotechnology. Insects are one of the most versatile groups of the animal kingdom. Due to their large population sizes and adaptability since long they attract researchers’ interest as efficient resource for agricultural and biotechnological purposes. Several economically important insects such as Silkworms, Honey Bee, Lac and Drosophila or Termites were established as invertebrate model organisms. Starting with the era of genetic engineering, a broad range of molecular and genetic tools have been developed to study the molecular biology of these insects in detail and thus opened up a new horizon for multidisciplinary research. Nowadays, insect derived products are widely used in biomedical and biotechnology industries. The book targets researchers from both academia and industry, professors and graduate students working in molecular biology, biotechnology and entomology.
Author: Elena García-Fruitós
Publisher: Humana Press
ISBN: 9781493922048
Category : Science
Languages : en
Pages : 0
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Book Description
With insolubility proving to be one of the most crippling bottlenecks in the protein production and purification process, this volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Insoluble Proteins: Methods and Protocols includes chapters that describe not only the recombinant protein production in different expression systems but also different purification and characterization methods to finally obtain these difficult-to-obtain proteins. Beginning with protein production methods using both prokaryotic and eukaryotic expression systems, the book continues with purification protocols using insoluble proteins, the characterization of insoluble proteins, as well as a general overview of interesting applications of insoluble proteins. 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. Comprehensive and practical, Insoluble Proteins: Methods and Protocols aims to provide the scientific community with detailed and reliable state-of-the-art protocols that are used in order to successfully produce and purify recombinant proteins prone to aggregate.
Author: Joseph M. Fernandez
Publisher: Elsevier
ISBN: 0080532357
Category : Science
Languages : en
Pages : 493
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Book Description
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: Michael R. Dyson
Publisher: Methods Express (Hardcover)
ISBN:
Category : Science
Languages : en
Pages : 318
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Book Description
Protein expression is an increasingly important tool for research on gene function. What is needed is not just a lab manual providing established methods as well as the latest state-of-the-art protocols, but also clear advice on what expression system to choose when. Expression Systems: Methods Expressuniquely fills this need. It covers expression across a broad range of systems, including the following. *Baculovirus expression vectors *CHO cells *E. coli *HEK293-EBNA1 cells *Lactococcus lactis and other gram positive bacteria *S. cerevisiae *transfected insect cells *Pichia pastoris *mammalian cells using BacMam viruses *lentiviral vectors *wheat germ cell-free system The book takes the reader through how to make an informed choice of appropriate system, taking into account the protein target, the time involved, the ultimate use of the expressed protein, and the laboratory equipment required. It also provides step-by-step methods for each system. In addition, the book describes the optimisation of expression strategies, expression engineering using ribosome display, and how to select protein variants with improved expression. Every chapter discusses the merits and limitations of the approaches available, describes the key techniques in full practical detail, and provides sensible advice for immediate use at the bench. In summary, Expression Systems: Methods Expressis a comprehensive laboratory manual and information resource for researchers at all levels, from postgraduate student to principal investigator.
