Overview of the Baculovirus Expression System

Cheryl Isaac Murphy1, Helen Piwnica‐Worms2

1 Cambridge Biotech Corporation, Worcester, Massachusetts, 2 Washington University School of Medicine, St.Louis, Missouri
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 4.18
DOI:  10.1002/0471142301.ns0418s10
Online Posting Date:  May, 2001
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Baculoviruses have emerged as a popular system for overproducing recombinant proteins in eukaryotic cells. This unit gives an overview of the baculovirus expression system, including discussion of the baculovirus life cycle, and postā€translational modifications that occur in insect cells. In addition, the steps for overproducing proteins in the baculovirus systems are described along with recommendations for choosing an appropriate baculovirus vector and DNA, and reagents and equipment necessary for implementing the whole overexpression system.

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Table of Contents

  • Baculovirus Life Cycle
  • Baculovirus Expression System
  • Post‐Translational Modification of Proteins in Insect Cells
  • Figures
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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 pp60c‐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.
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