Skip to main content

Baculovirus Expression Vectors

  • Protocol
Practical Molecular Virology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 8))

Abstract

The baculovirus system has the potential for the large-scale production of protein products by two methods. First, as a result of recent advances (1,5), large-scale cell culture is now possible and, second, the recombinant baculovirus can be used to infect susceptible insect larvae (3). Whether using small-scale cell culture or the aforementioned methods for scale-up, the requirement for downstream processing of the protein product is manifest. Purification of the product is greatly facilitated when expression is high (ca. 30% of total cell protein); however, when expression is relatively low or membrane proteins are required, the researcher faces the same problems encountered in ascertaining the optimum conditions for any de novo protein purification. Obviously, the degree of purity required for a given product is indicative of its final use; consequently, a number of antigens for use in diagnostic procedures have been relatively crude preparations (4,5). This chapter highlights the methods that have been used to purify baculovirusexpressed protein products, and is aimed at providing general guidelines to the purification of certain types of protein product rather than a definitive guide to protein purification.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Maiorella, B., Inlow, D., Shauger, A., and Harano, D. (1988) Large scale msect cell culture for recombinant protein production. Bio/Technology l6, 1406–1410.

    Article  Google Scholar 

  2. Murhammer, D. W. and Coochee, C. F. (1988) Scaleup of insect cell cultures Protective effects of Pleromc F-68. Bio/Technology 6, 1411–1418.

    Article  CAS  Google Scholar 

  3. Emery, V. C. and Bishop, D. H. L. (1987) The development of multtple expression vectors for the high level synthesis of eukatyotic proteins: Expression of theLCMV-N and AcNPV polyhedrin protein by a recombinant baculovirus. Protein Eng. 1, 393–366.

    Article  Google Scholar 

  4. Madtsen, L., Travis, B., Hu, S L, and Purchio, A. F. (1987) Expression of the human immunodeficiency virus gag gene m insect cells. Virology~5 8,248-250.

    Google Scholar 

  5. Schmaljohn, C. S., Sugiyama, R., Schmaljohn, A. L., and Bishop, D. H. L. (1988) Baculovirus expression of the small genome segment of Hantaan virus and potential use of the expressed nucleocapsid protein as a diagnostic antigen. J Gcn. Viral, 69, 777-786.

    Google Scholar 

  6. Hoffman, J. D., Lawson, F. R., and Yamamoto, R. (1966) Synthetic growth media for propagating insect larvae, m InsecCt olonbztwna ndM msProductm (Smith, C. N., ed.), Academic, NY, pp. 155-190.

    Google Scholar 

  7. Takeham, K., Ireland, D., and Bishop, D. H. L., (1988) &expression of the Hepatitis B surface and core antigens using baculovirus multiple expression vectors.J. Cen viral 69,2763-2777.

    Google Scholar 

  8. Ray, R., Galinski, M. S., and Compans, R. W. (1989) Expression of the fusion glycop rotem of human parainfluenza type 3 virus in insect cells by a recombinant baculovirus and analysis of its immunogenicity. Vinu. Bcs. 12,169-180.

    Google Scholar 

  9. Wedegaertner, P. B. and Gill, G. N. (1989) Activation of the purified protein kinase domain of the epidennal growth factor receptor. J. BwL Chm. 264,1134611353.

    Google Scholar 

  10. Cuatrecasas, P. (1970) Protein purification by afftnity chromatography.J. BioL Chem. 245,3059-3065.

    Google Scholar 

  11. Webb, N. R., Madoulet, C., Tosi, P.F., Broussard, D. R., Sneed, L., Nicolau, C., and Summers, M. D. (1989) Cell surface expression and purification of human CD4 produced in baculovirus infected cells. Pm. Nail. Acad. So USA 86,7731-7735.

    Google Scholar 

  12. Price, P. M., Reichelderfer, C. F., Johansson, B. E., Rilboume, E. D., and Acs, G (1989) Complementadon of recombinan t baculovlruses by comfec tion with wild typevu-us facrhtates producuon in msect larvae of antigenic proteins of hepautis B vrrus and mfluenza virus. Proc. NatL Acad. Sea. USA 86,1453-1457.

    Google Scholar 

  13. Bishop, D. H. L. and Emery, V. C. (1988) Expression vectors for the synthesis of protems and plasmtd replicons and sequence cassettes for use in constructing such vectors. Eur. Patent 88807452.8.

    Google Scholar 

  14. Bishop, D. H. L. and Emery, V. C. (1988) Expression vectors for the synthesis of proteins and plasmid replicons and sequence cassettes for use in constructing such vectors US Patent PCT/GB88/00663.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1991 The Humana Press Inc.

About this protocol

Cite this protocol

Emery, V.C. (1991). Baculovirus Expression Vectors. In: Collins, M.K.L. (eds) Practical Molecular Virology. Methods in Molecular Biology, vol 8. Humana Press. https://doi.org/10.1385/0-89603-191-8:319

Download citation

  • DOI: https://doi.org/10.1385/0-89603-191-8:319

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-191-3

  • Online ISBN: 978-1-59259-495-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics