Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette

PLoS One. 2014 May 13;9(5):e96562. doi: 10.1371/journal.pone.0096562. eCollection 2014.

Abstract

Here we describe the development of a baculovirus vector expression cassette containing rearranged baculovirus-derived genetic regulatory elements. This newly designed expression cassette conferred significant production improvements to the baculovirus expression vector system (BEVS), including prolonged cell integrity after infection, improved protein integrity, and around 4-fold increase in recombinant protein production yields in insect cells with respect to a standard baculovirus vector. The expression cassette consisted of a cDNA encoding for the baculovirus transactivation factors IE1 and IE0, expressed under the control of the polyhedrin promoter, and a homologous repeated transcription enhancer sequence operatively cis-linked to the p10 promoter or to chimeric promoters containing p10. The prolonged cell integrity observed in cells infected by baculoviruses harbouring the novel expression cassette reduced the characteristic proteolysis and aberrant forms frequently found in baculovirus-derived recombinant proteins. The new expression cassette developed here has the potential to significantly improve the productivity of the BEVS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cell Line
  • Gene Expression
  • Genetic Vectors*
  • Insecta / genetics
  • Promoter Regions, Genetic*
  • Recombinant Proteins / genetics*

Substances

  • Recombinant Proteins

Grants and funding

This work was supported by the grant CSD2006-00007-2 and RTA2010-00084-C02-01 from the Spanish Ministry of Economy and Competitiveness and by funds from the company Alternative Gene Expression S.L. (ALGENEX). Alternative Gene Expression provided support in the form of salaries for authors SGV and JLV but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the “author contributions” section.