Baculovirus Expression Vectors: A Laboratory Manual

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Oxford University Press, 1994 - Science - 347 pages
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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 appropriate baculovirus 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; guidelines for 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 comprehensive approach 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. The manual is an invaluable resource for every individual engaged in the production of proteins for any purpose.
 

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Contents

Virus Structure and the Infection Process
3
Gene Organization Regulation and Function
12
VirusHost Interactions
24
CHOOSING A TRANSFER PLASMID AND PARENT VIRUS
33
Choosing a Parent Virus for Use in Vector Construction
94
METHODS FOR VECTOR CONSTRUCTION
105
Virus Methods
124
Cotransfection and Recombinant Virus Identification
139
Recommended Operating Conditions
244
Running the Bioreactor
253
Nucleotide Sequence of AcMNPV
271
Nucleotide Sequence of AcMNPV plO Region
280
List of Genes Expressed in the Baculovirus Expression System
285
Suppliers
297
Stock Solutions
302
Excel Spreadsheet for TCIDgo Calculation
304

Characterizing Recombinant Gene Expression
180
PostTranslational Modification
216
Production in Cell Culture versus Insect Larvae
237
References
307
Index
341
Copyright

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Page 335 - Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.

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About the author (1994)

David R. O'Reilly is at Imperial College, London. Lois K. Miller is at University of Georgia.

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