Baculovirus and Insect Cell Expression Protocols
Baculovirus and Insect Cell Expression Protocols

Author: David W. Murhammer

Publisher: Methods in Molecular Biology

Published: 2018-04-15

Total Pages: 522

ISBN-13: 9781493979783

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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.

Baculovirus and Insect Cell Expression Protocols
Baculovirus and Insect Cell Expression Protocols

Author: David W. Murhammer

Publisher: Humana Press

Published: 2007-07-06

Total Pages: 470

ISBN-13: 9781588295378

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Baculovirus Expression Protocols, Second Edition, provides the detailed steps required to perform the techniques involved with the use of baculoviruses and insect cell culture and discusses problems that may be encountered. This newly-revised and updated edition provides a cutting edge step-by-step guide for biochemists, molecular biologists, biochemical engineers and others using the BEVS and/or insect cells for producing recombinant proteins. Furthermore, Baculovirus Expression Protocols, Second Edition, provides assistance to scientists and engineers interested in developing and producing baculovirus insecticides. Authors list all available insect cell lines and provide methods for isolating new cell lines. The procedures involved in producing products, both at laboratory scale and large scale, are discussed, as well as the production in insect larvae. It is hoped that this volume will not only aid the user in successfully completing the tasks described herein, but will also stimulate the development of improved techniques and new applications of baculoviruses and insect cell culture. Timely and thorough, this volume will become an essential reference for anyone using baculoviruses and insect cell culture.

Recombinant Protein Expression: Eukaryotic hosts
Recombinant Protein Expression: Eukaryotic hosts

Author:

Publisher: Academic Press

Published: 2021-11-04

Total Pages: 384

ISBN-13: 0323907385

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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

Baculovirus Expression Vectors
Baculovirus Expression Vectors

Author: David R. O'Reilly

Publisher:

Published: 1994

Total Pages: 366

ISBN-13: 9780195091311

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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.

Baculovirus Expression Protocols
Baculovirus Expression Protocols

Author: Christopher D. Richardson

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 412

ISBN-13: 159259526X

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Baculovirus Expression Protocols offers both industrial and university-based researchers a comprehensive comilation of the latest baculovirus techniques along with step-by-step instructions and time-saving techniques. The contributors-leading authorities in the field-present the assorted expression plasmids currently in use, guide the reader through the process of generating and selecting recombinant virus, and describe specific examples of recombinant protein production and purification. The emphasis is on alternative and simpler screening techniques for the selection of recombinant baculovirus. The book also surveys the various insect cell lines currently compatible with the baculovirus system. Highlights include production of recombinant virus using linearized DNA and vectors that contain a b-galactosidase indicator and a complete list of expression vectors currently available. Detailed descriptions for the scale-up of protein production using spinner flasks, bioreactors, and insect larvae are also included. Baculovirus Expression Protocols offers both industrial and university-based researchers an outstanding collection of reproducible, step-by-step laboratory protocols. It will immediately become indispensable for anyone working with baculoviruses and their application in the expression of recombinant proteins in insect cells.

Viral Vectors for Gene Therapy
Viral Vectors for Gene Therapy

Author: Fredric P. Manfredsson

Publisher:

Published: 2019

Total Pages: 328

ISBN-13: 9781493990658

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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.

Trends in Insect Molecular Biology and Biotechnology
Trends in Insect Molecular Biology and Biotechnology

Author: Dhiraj Kumar

Publisher: Springer

Published: 2018-04-03

Total Pages: 376

ISBN-13: 331961343X

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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.

Advanced Technologies for Protein Complex Production and Characterization
Advanced Technologies for Protein Complex Production and Characterization

Author: M. Cristina Vega

Publisher: Springer

Published: 2016-05-10

Total Pages: 378

ISBN-13: 3319272160

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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.

Protein Chromatography
Protein Chromatography

Author: Dermot Walls

Publisher: Humana Press

Published: 2011-04-06

Total Pages: 527

ISBN-13: 9781607619147

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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.

The Baculoviruses
The Baculoviruses

Author: Lois K. Miller

Publisher: Springer Science & Business Media

Published: 1997-07-31

Total Pages: 480

ISBN-13: 9780306456411

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The past decade has witnessed an explosion of information on the molecular biology of insect viruses and a frenzy of activity in applying this information to medicine and agriculture. Genetically engineered baculoviruses are presently being tested for commercial use as pesticides, and the study of such viruses is also revealing remarkable insights into basic cellular processes such as apoptosis. This comprehensive volume provides readers with knowledge of basic and applied baculovirology so that current literature in the field can be appreciated.