1 The baculoviruses -- 1.1 Introduction -- 1.2 Isolation and host range -- 1.3 Structure and classification -- 1.4 Baculovirus replication in vivo -- 1.5 Baculovirus replication in vitro -- 1.6 Genetic engineering of baculovirus insecticides -- 2 The development of baculovirus expression vectors -- 2.1 Introduction and historical perspective -- 2.2 The merits of the baculovirus expression system -- 2.3 General principles for inserting foreign genes into the baculovirus genome -- 2.4 Baculovirus transfer vectors -- 2.5 Selection of recombinant viruses -- 3 Processing of foreign proteins synthesized using baculovirus vectors in insect cells -- 3.1 Introduction -- 3.2 Glycosylation -- 3.3 Phosphorylation, acylation and amidation -- 3.4 Proteolytic processing -- 3.5 Cellular targeting and secretion -- 3.6 Tertiary and quaternary structure formation -- 3.7 Expression of viral genes -- 3.8 Expression of bacterial and fungal genes -- 3.9 Post-transcriptional processing -- 4 Construction of transfer vectors containing the foreign gene -- 4.1 Introduction -- 4.2 Isolation of foreign gene coding sequences -- 4.3 Modifying the ends of DNA molecules -- 4.4 Preparation of the transfer vector -- 4.5 DNA ligations -- 4.6 Transformation of bacteria -- 4.7 Screening for recombinant baculovirus transfer vectors -- 4.8 Analysis of recombinant transfer vectors -- 4.9 Isolation of highly purified plasmid DNA (maxi-preps) -- 5 Insect cell culture media and maintenance of insect cell lines -- 5.1 Introduction -- 5.2 Cell lines -- 5.3 Culture media -- 5.4 Preparation of culture media -- 5.5 Glassware and disposable plasticware -- 5.6 Insect cell culture -- 5.7 A guide to Sf cell seeding densities for experimental work -- 5.8 Freezing, storage and recovery of insect cells in liquid nitrogen -- 5.9 A guide to adapting cells to serum-free media -- 6 Propagation, titration and purification of AcMNPV in cell culture -- 6.1 Introduction -- 6.2 Infection of cells with virus for experimental work -- 6.3 Titration of virus by plaque-assay -- 6.4 Plaque-picking and plaque-purification -- 6.5 Amplification of virus stocks -- 6.6 Large-scale production of virus for the purification of virus particles -- 6.7 Purification of infectious virus DNA -- 6.8 Titration of virus by TCID50 -- 7 Production and selection of recombinant virus -- 7.1 Introduction -- 7.2 Preparation of linear AcMNPV.lacZ (or AcMNPV.SC) DNA -- 7.3 Co-transfection of insect cells -- 7.4 Separation of parental and recombinant viruses by plaque-assay -- 7.5 Plaque-purification and amplification of recombinant virus stocks -- 7.6 Amplification and detection of recombinant viruses by limiting dilution and dot-blot hybridization -- 8 Characterization of recombinant viruses -- 8.1 Introduction -- 8.2 Analysis of recombinant virus genomes -- 8.3 Analysis of foreign gene expression by polyacrylamide gel electophoresis, using unlabelled or radiolabelled cell proteins -- 8.4 Analysis of recombinant protein synthesis in insect cells using immunological techniques -- 8.5 Analysis of post-translational processing events in insect cells -- 8.6 Analysis of transcription in recombinant virus-infected cells -- 9 Scaling up the production of recombinant protein in insect cells; laboratory bench level -- 9.1 Introduction -- 9.2 Large-scale culture of insect cells -- 9.3 The importance of highly infectious virus stocks -- 9.4 Multiplicity of infection -- 9.5 The optimum time to harvest virus-infected cells -- 9.6 Purification of recombinant protein from infected cell cultures -- 10 Propagation of baculoviruses in insect larvae -- 10.1 Introduction -- 10.2 Rearing insects in the laboratory -- 10.3 Infection of insect larvae with polyhedra from cell culture -- 10.4 Purification of polyhedra from infected larvae -- 10.5 Bioassays of polyhedra -- 10.6 Purification of virus particles and DNA from polyhedra -- 10.7 Isolation of virus particles from infected larvae to establish infections in cell culture -- 10.8 Preparation of semi-synthetic insect diet -- 11 Trouble-shooting guide -- 11.1 Introduction -- 11.2 Insertion of foreign gene coding sequences into transfer vectors -- 11.3 Cell culture -- 11.4 Preparation of virus stocks and infectious DNA -- 11.5 Co-transfections -- 11.6 Baculovirus plaque-assays -- 11.7 Screening for recombinant viruses -- 11.8 Instability of recombinant viruses -- 11.9 Poor yields of recombinant protein -- Appendix A list of selected suppliers -- References.