Recombinant Protein Expression in the Baculovirus-Infected Insect Cell System

  • Tamar UngerEmail author
  • Yoav Peleg
Part of the Methods in Molecular Biology book series (MIMB, volume 800)


Over the last two decades, the use of eukaryotic cells for expression of recombinant proteins has become the preferred choice for many applications. This is primarily the case when posttranslational modifications and correct disulfide-bond formation are necessary for protein folding and activity. Among the eukaryotic expression systems, the baculovirus-infected insect cell platform has gained particular attention, resulting in the development and implementation of multiple strategies for protein expression. Here, we present baculovirus-infected insect cells as an efficient expression system for eukaryotic proteins. We demonstrate a simplified and a shortened procedure for recombinant virus production that is sufficient for large-scale production of proteins in insect cells.

Key words

Baculovirus Insect cells Protein expression and production Secreted proteins Intracellular proteins 



We thank Prof. J. Sussman, Prof. I. Silman, Prof. G. Schreiber and Prof. Yigal Burstein for their continuous support throughout the study. This research was supported by the European Commission Sixth Framework Research and Technological Development Program ‘SPINE2-COMPLEXES’ Project, under contract No. 031220; a grant from the Israel Ministry of Science, Culture, and Sport to the Israel Structural Proteomics Center; the Divadol Foundation; and the Neuman Foundation.


  1. 1.
    Posse RD (1997) Baculoviruses as expression vectors. Curr Opin Biotechnol 8: 569–572CrossRefGoogle Scholar
  2. 2.
    Luckow VA, and Summers MD (1988) Signals important for high-level expression of foreign genes in Autographa californica nuclear polyhedrosis virus expression vectors. Virology 167: 56–71PubMedCrossRefGoogle Scholar
  3. 3.
    Fraser MJ (1986) Ultrastructural observations of virion maturation in Autographa californica Nuclear Polyhedrosis virus infected Spodoptera frugiperda cell cultures. J Ultrastruct Mol Struct Res 95: 189–195CrossRefGoogle Scholar
  4. 4.
    Summers MD, Anderson DL (1972) Characterization of deoxyribonucleic acid isolated from the granulosis viruses of the cabbage looper, Trichoplusia ni and the fall armyworm, Spodoptera frugiperda. Virology 50: 459-471PubMedCrossRefGoogle Scholar
  5. 5.
    Kost TA, Condreay JP, Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 23: 567–575PubMedCrossRefGoogle Scholar
  6. 6.
    Chen X, Liu H, Shim AH, Focia PJ, He X (2008) Structural basis for synaptic adhesion mediated by neuroligin-neurexin interactions. Nat Struct Mol Biol 15: 50–56PubMedCrossRefGoogle Scholar
  7. 7.
    Sun XZ, Nguyen J, Momand J (2003) Purification of recombinant p53 from Sf9 insect cells. Methods Mol Biol 234: 17–28PubMedGoogle Scholar
  8. 8.
    Jarvis DL (2009) Baculovirus-insect cell expression systems. Methods Enzymol 463: 191–222PubMedCrossRefGoogle Scholar
  9. 9.
    Luckow VA, Lee SC, Barry GF, Olins PO (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: 4566–4579PubMedGoogle Scholar
  10. 10.
    Vialard JE, Richardson CD (1993) The 1,629-nucleotide open reading frame located downstream of the Autographa californica nuclear polyhedrosis virus polyhedrin gene encodes a nucleocapsid-associated phosphoprotein. J Virol 67: 5859–5866PubMedGoogle Scholar
  11. 11.
    Invitrogen (2002) Growth and maintenance of insect cell lines. Invitrogen Life Technologies, version K, Paisley, UKGoogle Scholar
  12. 12.
    Shrestha B, Smee C, Gileadi O (2008) Baculovirus expression vector system: an emerging host for high-throughput eukaryotic protein expression. Methods Mol Biol 439P: 269–289PubMedCrossRefGoogle Scholar
  13. 13.
    Pennock GD, Shoemaker C, Miller LK (1984) Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector. Mol Cell Biol 4: 399–406PubMedGoogle Scholar
  14. 14.
    van den Ent F, Lowe J (2006) RF cloning: a restriction-free method for inserting target genes into plasmids. J Biochem Biophys Methods 67: 67–74PubMedCrossRefGoogle Scholar
  15. 15.
    Unger T, Jacobovitch Y, Dantes A, Bernheim R, Peleg Y (2010) Applications of the Restriction Free (RF) cloning procedure for molecular manipulations and protein expression. J Struct Biol 172: 34–44PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Israel Structural Proteomics Center, Faculty of BiochemistryWeizmann Institute of ScienceRehovotIsrael

Personalised recommendations