Baculovirus and Insect Cell Expression Protocols

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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Contenu

Useful Tips, Widely Used Techniques And Quantifying Cell Metabolic Behavior.- Introduction to Baculovirus Molecular Biology.- Baculovirus Transfer Vectors.- Recombinant Baculovirus Isolation.- Gene Expression In Mammalian Cells Using Bacmam, A Modified Baculovirus System.- Available Lepidopteran Insect Cell Lines.- Lepidopteran Insect Cell Line Isolation from Insect Tissue.- Development Of Serum-Free Media For Lepidopteran Insect Cell Lines.- Routine Maintenance And Storage Of Lepidopteran Insect Cell.- Small-Scale Production Of Recombinant Proteins Using The Baculovirus Expression Vector System.- Recombinant Protein Production In Large Scale Agitated Bioreactors Using The Baculovirus Expression Vector System.- Protein Expression in Insect and Mammalian Cells Using Baculoviruses in Wave Bioreactors.- Protein Production With Recombinant Baculoviruses in Lepidopteran Larvae.- Production Of Virus-Like Particles For Vaccination.- Alternative Strategies for Expressing Multi-Component Protein Complexes in Insect Cells.- Transforming Lepidopteran Insect Cells for Continuous Recombinant Protein Expression.- Stable Drosophila Cell Lines: An Alternative Approach to Exogenous Protein Expression.- Transforming Lepidopteran Insect Cells for Improved Protein Processing and Expression.- Introduction To The Use Of Baculoviruses As Biological Insecticides.- Baculovirus Insecticide Production In Insect Larvae.- Evaluation Of The Insecticidal Efficacy Of Wild Type And Recombinant Baculoviruses.- Evaluating Baculovirus Infection Using Green Fluorescent Protein and Variants.- Tubular Bioreactor for Probing Baculovirus Infection and Protein Production.- Gene Silencing In Insect Cells Using Rnai.- Using the Baculovirus/Insect Cell System to Study Apoptosis.- Generation of Envelope-Modified Baculoviruses for Gene Delivery into Mammalian Cells.