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Overview of the Baculovirus Expression System

Cheryl Isaac Murphy¹, Helen Piwnica‐Worms²

¹Cambridge Biotech Corporation, Worcester, Massachusetts
²Washington University School of Medicine, St. Louis, Missouri

Publication Name:

Current Protocols in Protein Science

Unit Number:

Unit 5.4

DOI:

10.1002/0471140864.ps0504s00

Online Posting Date:

May, 2001

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Abstract

Baculoviruses have emerged as a popular system for overproducing recombinant proteins in eukaryotic cells. This overview unit describes the baculovirus life cycle and expression system, and also provides information on vectors and protocols for using the baculovirus expression system.

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Unit Introduction
Baculovirus Life Cycle
Baculovirus Expression System
Post-Translational Modification of Proteins in Insect Cells
Bibliography
Figures

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Figures

Figure 5.4.1

Baculovirus life cycle. Viruses enter cells by adsorptive endocytosis and move to the nucleus where their DNA is released. Both DNA replication and viral assembly take place in the nuclei of infected cells to generate two types of viral progeny. These include extracellular (nonoccluded) virus particles and polyhedra-derived (occluded) virus particles. Extracellular virus is released from the cell by budding, starting at ~12 hr postinfection and ending ~36 hr postinfection. Polyhedra-derived virus appears later (~18 hr postinfection) and accumulates in the nuclei of infected cells £72 hr postinfection or until cellular lysis. Polyhedra-derived virus is embedded in proteinaceous viral occlusions, the major protein component of which is the viral polyhedrin protein. Secondary infection of cells and tissues occurs by two pathways. In the first, the extracellular virus, once budded from the site of primary infection, is free to infect neighboring cells by the pathway just described. Alternatively, polyhedra-derived virus is released from occlusion bodies after an infected food source is ingested by a new host. Reproduced from Summers and Smith (1987) with permission from the Texas Agricultural Experiment Station.

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Literature Cited

Literature Cited
	Carbonell, L.F., Klowden, M.J., and Miller, L.K. 1985. Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells. J. Virol. 56:153-160.
	Doerfler, W. and Bohm, P. 1986. The Molecular Biology of Baculoviruses. Springer-Verlag, New York.
	Hellers, M., Gunne, H., and Steiner, H. 1991. Expression and post-translational processing of preprocecropin-A using a baculovirus vector. Eur. J. Biochem. 199:435-439.
	Jarvis, D.L. and Summers, M.D. 1992. Baculovirus expression vectors. In Recombinant DNA Vaccines: Rationale and Strategies (R.E. Isaacson, ed.) pp.265-291. Marcel Dekker, New York.
	Kitts, P.A. and Possee, R.D. 1993. A method for producing recombinant baculovirus expression vectors at high frequency. BioTechniques 14:810-817.
	Kitts, P.A., Ayres, M.D., and Possee, R.D. 1990. Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucl. Acids Res. 18:5667-5672.
	Luckow, V.A. 1991. Cloning and expression of heterologous genes in insect cells with baculovirus vectors. In Recombinant DNA Technology and Applications (A. Prokop, R.K. Bajpai, and C. Ho, eds.) pp.97-152. McGraw-Hill, New York.
	Luckow, V.A. and Summers, M.D. 1988. Trends in the development of baculoviral expression vectors. Bio/Technology 6:47-55.
	Miller, L.K. 1988. Baculoviruses as gene expression vectors. Annu. Rev. Microbiol. 42:177-199.
	Miller, D.W., Safer, P., and Miller, L.K. 1986. An insect baculovirus host vector for high-level expression of foreign genes. In Genetic Engineering, Vol. 8 (J.K. Setlow and A. Hollaender eds.) pp.277-298. Plenum, New York.
	O'Reilly, D.R., Miller, L.K., and Luckow, V.A. 1992. Baculovirus Expression Vectors. W.H. Freeman and Company, New York.
	Piwnica-Worms, H., Williams, N.G., Cheng, S.H., and Roberts, T.M. 1990. Regulation of pp60^c-src and its association with polyoma virus middle T antigen in insect cells. J. Virol. 64:61-68.
	Summers, M.D. and Smith, G.E. 1987. A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin No. 1555. College Station, Texas.
Key Reference
	O'Reilly et al. 1992. See above.
A guide assembled to aid researchers using the baculoviral expression system, containing detailed protocols for using this system effectively.