Baculovirus Insect Cell Expression System

Abstract

The baculovirus insect cell expression system is based on the use of recombinant baculoviruses (insect viruses) and their ability to produce high levels of biologically active proteins in cultured insect cells or insect larvae. These viruses have very late genes (polyhedrin, P10), which are highly expressed due to very strong promoters, but are dispensable for infectious virus formation. Deletion of these genes and replacement with foreign genes convert these viruses into expression vectors for the production of recombinant proteins. Convenient systems (bacmids) are available for engineering these baculovirus recombinants in bacteria prior to transfection into insect cells. The proteins produced can be easily purified from infected cells or their supernatants using tags and affinity chromatography. The proteins can be used for human and veterinary applications, such as vaccines and diagnostics.

Keywords: eukaryotic expression systems; insect cell technology; virology; diagnostics; subunit vaccines

Figure 1.

Baculovirus insect cell expression system. In cells infected with a wild‐type virus (a) large amounts of polyhedra and p10 are formed. In most cases in the recombinant virus (b) the polyhedrin and/or p10 promoter is used to drive the expression of a foreign gene and the gene cassette replaces the polyhedrin gene, resulting in a recombinant with a polyhedron‐negative phenotype and (c) high levels of recombinant protein are produced in stead of polyhedrin (lanes 1 and 2).

close

References

Harrison RL and Jarvis DL (2006) Protein N‐glycosylation in the baculovirus‐insect cell expression system and engineering of insect cells to produce “mammalianized” recombinant glycoproteins. Advances in Virus Research 68: 159–191.

Harro CD, Pang YY, Roden RB et al. (2001) Safety and immunogenicity trial in adult volunteers of a human papillomavirus 16 L1 virus‐like particle vaccine. Journal of the National Cancer Institute 93: 284–292.

Hulst MM, Westra DF, Wensvoort G and Moorman RJ (1993) Glycoprotein E1 of hog cholera virus expressed in insect cells protects swine from hog cholera. Journal of Virology 67: 5435–5442.

Kaba SA, Salcedo AM, Wafula PO, Vlak JM and van Oers MM (2004) Development of a chitinase and v‐cathepsin negative bacmid for improved integrity of secreted recombinant proteins. Journal of Virological Methods 122: 113–118.

Kitts PA, Ayres MD and Possee RD (1990) Linearization of baculovirus DNA enhances the recovery of recombinant virus expression vectors. Nucleic Acids Research 18: 5667–5672.

Luckow VA, Lee SC, Barry GF and Olins PO (1993) Efficient generation of infectious recombinant baculoviruses by site‐specific transposon‐mediated insertion of foreign genes into a baculovirus genome propagated in E. coli. Journal of Virology 67: 4566–4579.

Martens JW, van Oers MM, van de Bilt BD, Oudshoorn P and Vlak JM (1995) Development of a baculovirus vector that facilitates the generation of p10‐based recombinants. Journal of Virological Methods 52: 15–19.

Pijlman GP, de Vrij J, van den End FJ, Vlak JM and Martens DE (2004) Evaluation of baculovirus expression vectors with enhanced stability in continuous cascaded insect‐cell bioreactors. Biotechnology and Bioengineering 87: 743–753.

Pijlman GP, Roode EC, Fan X et al. (2006) Stabilized baculovirus vector expressing a heterologous gene and GP64 from a single bicistronic transcript. Journal of Biotechnology 123: 13–21.

Possee RD, Thomas CJ and King LA (1999) The use of baculovirus vectors for the production of membrane proteins in insect cells. Biochemical Society Transactions 27: 928–932.

Smith GE, Summers MD and Fraser MJ (1983) Production of human beta interferon in insect cells infected with a baculovirus expression vector. Molecular and Cellular Biology 3: 2156–2165.

Summers MD (2006) Milestones leading to the genetic engineering of baculoviruses as expression vector systems and viral pesticides. Advances in Virus Research 68: 3–73.

Theilmann DA, Blissard GW, Bonning B et al. (2005) Family Baculoviridae. In: Fauquet CM, Mayo MA, Maniloff J, Desselberger U and Ball LA (eds) Virus Taxonomy, VIIIth Report of the ICTV, 8th edn, pp. 177–185. London: Elsevier/Academic Press.

Van Oirschot JT (1999) Diva vaccines that reduce virus transmission. Journal of Biotechnology 73: 195–205.

Further Reading

Condreay JP and Kost TA (2007) Baculovirus expression vectors for insect and mammalian cells. Current Drug Targets 8: 1126–1131.

Hu YC (2006) Baculovirus vectors for gene therapy. Advances in Virus Research 68: 287–320.

van Oers MM (2006) Vaccines for viral and parasitic diseases produced with baculovirus vectors. Advances in Virus Research 68: 193–253.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
van Oers, Monique M, and Vlak, Just M(Dec 2008) Baculovirus Insect Cell Expression System. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002660.pub2]