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

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

Author information

1
Alternative Gene Expression S.L. (ALGENEX), Madrid, Spain.
2
Department of Biotechnology, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.

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.

PMID:
24824596
PMCID:
PMC4019511
DOI:
10.1371/journal.pone.0096562
[Indexed for MEDLINE]
Free PMC Article

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