The Baculoviruses

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.

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.

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.

V.1 - Biological properties and molecular biology: Historical perspectives; Taxonomy and identification of baculoviruses; Ultrastructure of baculoviruses; In vivo infection and replication of baculoviruses; In vitro replication of baculoviruses; Baculovirus structural proteins and protein synthesis; Current developments in baculovirus serology; Persistent baculovirus infections; Specificity and safety of baculoviruses; Evolution of occluded baculoviruses; The genetics of baculoviruses; Organization and expression of the baculovirus genome; v.2 - Practical application for insect control: In vivo and in vitro bioassay methods for baculoviruses; In vivo production of baculoviruses; Cell culture methods for large-scale propagation of baculoviruses; Ecology and ens, chemical pesticides, and parasitoids; Formulation and application of baculoviruses; Use of baculoviruses in pest management programs; Registration of baculoviruses as pesticides; Practical factors influencing the utilization of baculoviruses as pesticides; Insect resistance to baculoviruses.

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Baculoviruses are perhaps unique among viruses in the breadth of their biotechnological applications: these insect specific viruses are used not only for insect pest management purposes, but also as laboratory research tools for production of recombinant proteins and for protein display, and as potential vectors for human gene therapy. In addition to highlighting recent advances, this volume provides a comprehensive review of the biotechnological applications of these and other insect viruses in both the academic and private sectors.

Developing effective baculovirus-insect culture systems. Insect cell culture methods and their use in virus research. Comparison of mammalian and insect cell cultures. protein production and processing from baculovirus. Development and testing of genetically improved baculovirus insecticides. Fundamentals of baculovirus-insect cell attachment and infection. Development and evaluation of host insect cells. Bioreactor design and scale-up issues. The effect of hydrodynamic forces on insect cells. Commercial application of insect cell culture. Baculovirus-mediated production of proteins in insect cells. Potential application of insect cell-based expression systems in the bio/pharmaceutical industry.