Baculovirus Expression Vectors: Improving Virus-Infected Cell Viability and Recombinant Protein Secretion

  • R. D. Possee
  • L. McCarroll
  • C. J. Thomas
  • S. G. Mann
  • L. A. King
Part of the Cell Engineering book series (CEEN, volume 1)


Baculoviruses are arthropod-specific viruses with a genome of double-stranded DNA (90-160 kilobase pairs, kbp). No baculoviruses have been described that replicate in vertebrates or plants. The viruses have to be propagated in arthropod larvae or in arthropod cells. Baculoviruses have been developed as successful expression vectors for the following reasons: (a) baculoviruses are easy to work with and safe to handle, (b) recombinant baculoviruses are genetically stable, (c) baculoviruses can be used as helper-independent vectors of foreign gene products, (d) large genes and cistrons can be expressed and (e) high levels of the desired expression products are often obtained. Although the system was originally established to enable the synthesis of individual, native gene products, a range of other expression vectors are now available. Some permit the production of recombinant proteins fused with various heterologous sequences to facilitate easy purification. Other vectors can be used to produce multiple proteins in insect cells. Recombinant products can also be incorporated as structural proteins into the budded form of the baculovirus particle. It is also proposed to use baculoviruses as biosafe vectors for human somatic gene therapy. The preparation of recombinant baculoviruses is straightforward and has benefited from the development of a number of novel procedures that enable the non-insect virologist to use the system with ease. Recent reviews provide an historical perspective of the development of the baculovirus expression systems and describe various features which are not summarised in this article (King and Possee, 1992; O’Reilly et al, 1992; Possee, 1997; Jarvis, 1997)


Insect Cell Recombinant Protein Production Chitinase Gene Nuclear Polyhedrosis Virus Insect Cell Culture 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ayres, M.D., Howard, S.C., Kuzio, J., Lopez-Ferber, M. and Possee, R.D. (1994) The complete DNA sequence of Autographa californiea nuclear polyhedrosis virus. Virology 202, 586–605.PubMedCrossRefGoogle Scholar
  2. Blissard, G.W. and Rohrmann, G.F. (1989) Location, sequence, transcriptional mapping and temporal expression of the gp64 envelope glycoprotein gene of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology 170, 537–542PubMedCrossRefGoogle Scholar
  3. Ernst, W.J., Grabherr, R.M. and Katinger, H.W. (1994) Direct cloning into the Autographa californica nuclear polyhedrosis virus for generation of recombinant baculoviruses. Nucleic Acids Res. 22, 2855–2857.PubMedCrossRefGoogle Scholar
  4. Friesen, P.D. (1997) Regulation of baculovirus early gene expression, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 141–170.Google Scholar
  5. Funk, C.J., Braunagel, S.C. and Rohrmann, G.F. (1997) Baculovirus structure, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 7–27.Google Scholar
  6. Guarino, L.A., Xu B., Jin, J and Dong, W. (1998) A virus-encoded RNA polymerase purified from baculovirus-infected cells. J. Virol. 72, 7985–7991.PubMedGoogle Scholar
  7. Hawtin, R.E., Arnold, K., Ayres, M.D., Zanotto, P.M de A., Howard, S.C, Gooday, G.A., Chappell, L.H., Kitts, P.A., King, L.A., and Possee, R.D. (1995) Identification and preliminary characterisation of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212, 673–685PubMedCrossRefGoogle Scholar
  8. Hawtin, R.E., Zarkowska, T., Arnold, K., Thomas, C.A., Gooday, G.W., King, L.A. and Possee, R.D. (1997) Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes. Virology 238, 243–253.PubMedCrossRefGoogle Scholar
  9. Jarvis, D.L. (1993) Effects of baculovirus infection on IE1-mediated foreign gene expression in stably transformed insect cells. J. Virol. 67, 2583–2588.PubMedGoogle Scholar
  10. Jarvis, D.L. (1997) Baculovirus expression vectors, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 389–431.Google Scholar
  11. King, L.A. and Possee, R.D. (1992) The Baculovirus Expression System: A Laboratory Guide, Chapman and Hall, London.Google Scholar
  12. Kitts, P.A. and Possee, R.D. (1993) A method for producing recombinant baculovirus expression vectors at high frequency. Biotechniques 14, 810–817.PubMedGoogle Scholar
  13. Lu, A. and Miller, L.K. (1997) Regulation of baculovirus late and very late gene expression, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 193–216.Google Scholar
  14. Lu, A., Krell, P.J., Vlak, J.M. and Rohrmann, G.F. (1997) Baculovirus DNA replication, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 171–191.Google Scholar
  15. Luckow, V.A., Lee, S.C., Barry, G.F. and Olins, P.O. (1993) Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J. Virol. 67, 4566–4571.Google Scholar
  16. O’Reilly, D.R. (1997) Auxiliary Genes of baculoviruses, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 267–300.Google Scholar
  17. Ohkawa, T., Majima, K. and Maeda, S. (1994) A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. J. Virol. 68, 6619–6625.PubMedGoogle Scholar
  18. O’Reilly, D.R., Miller, L.K. and Luckow, V.A. (1992) Baculovirus Expression Vectors, W.H. Freeman, New York.Google Scholar
  19. Possee, R.D. (1997) Baculoviruses as expression vectors. Current Opinion In Biotechnology 8, 569–572.PubMedCrossRefGoogle Scholar
  20. Rawlings, N.D., Pearl, L.H. and Buttle, D.F. (1992) The baculovirus Autographa californica multiple nuclear polyhedrosis virus genome includes a papain-like sequence. Biol. Chem. Hoppe-Seyler 373, 1211–1215.PubMedGoogle Scholar
  21. Slack, J.M., Kuzio, J. and Faulkner, P. (1995). Characterisation of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J. Gen. Virol. 76, 1091–1098.PubMedGoogle Scholar
  22. Suzuki, T., Kanaya, T., Okazaki, H., Ogawa, K., Usami, A., Watanabe, H., Kadono-Okuda, K., Yamakawa, M., Sato, H., Mori, H., Takahashi, S. and Oda, K. (1997) Efficient protein production using a Bombyx mori nuclear polyhedrosis virus lacking the cysteine proteinase gene. J. Gen. Virol. 78, 3073–3080.PubMedGoogle Scholar
  23. Thomas, C.J., Brown, H.L., Hawes, C.R., Lee, B. Y., Min, M.-K., Linda A. King, L.A. and Possee, R.D. (1998) Localisation of a baculovirus-induced chitinase in the insect cell endoplasmic reticulum. J. Virol. 72, 10207–10212.PubMedGoogle Scholar
  24. Volkman, L.E., Blissard, G.W., Friesen, P., Keddie, B.A., Possee, R.D. and Thielmann, D.A. (1995) Family Baculoviridae: In: Virus Taxonomy: Sixth Report of the International Committee on Taxonomy of Viruses, pp. 104–113. Edited by F.A. Murphy, C.M. Fauquet, D.H.L. Bishop, S.A. Ghabrial, A.W. Jarvis, G.P. Martelli, M.A. Mayo and M.D. Summers. Springer-Verlag Wien, New York.Google Scholar
  25. Whitford, M., Stewart, S., Kuzio, J. and Faulkner, P. (1989) Identification and sequence analysis of a gene encoding gp67 an abundant envelope glycoprotein of the baculovirus, Autographa californica nuclear polyhedrosis virus. J. Virol. 63, 1393–1398.PubMedGoogle Scholar
  26. Williams, G.V., Rohel, D.Z., Kuzio, J. and Faulkner, P. (1989) A cytopathological investigation of Autographa californica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants. J. Gen. Virol. 70, 187–202.PubMedCrossRefGoogle Scholar
  27. Williams, G.V. and Faulkner, P. (1997) Cytological changes and viral morphogenesis during baculovirus infection, in L.K. Miller (ed.), The Baculoviruses, Plenum Press, New York, pp. 61–107.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • R. D. Possee
    • 1
  • L. McCarroll
    • 2
  • C. J. Thomas
    • 2
  • S. G. Mann
    • 2
  • L. A. King
    • 2
  1. 1.NERC Institute of Virology and Environmental MicrobiologyOxfordUK
  2. 2.School of Biological and Molecular SciencesOxford Brookes UniversityUK

Personalised recommendations