Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette.
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 prol...
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doaj-91e9fc5749fd4473ad8af72b12633fca2020-11-25T02:33:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0195e9656210.1371/journal.pone.0096562Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette.Silvia Gómez-SebastiánJavier López-VidalJosé M EscribanoHere 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.http://europepmc.org/articles/PMC4019511?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Silvia Gómez-Sebastián Javier López-Vidal José M Escribano |
spellingShingle |
Silvia Gómez-Sebastián Javier López-Vidal José M Escribano Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. PLoS ONE |
author_facet |
Silvia Gómez-Sebastián Javier López-Vidal José M Escribano |
author_sort |
Silvia Gómez-Sebastián |
title |
Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
title_short |
Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
title_full |
Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
title_fullStr |
Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
title_full_unstemmed |
Significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
title_sort |
significant productivity improvement of the baculovirus expression vector system by engineering a novel expression cassette. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2014-01-01 |
description |
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. |
url |
http://europepmc.org/articles/PMC4019511?pdf=render |
work_keys_str_mv |
AT silviagomezsebastian significantproductivityimprovementofthebaculovirusexpressionvectorsystembyengineeringanovelexpressioncassette AT javierlopezvidal significantproductivityimprovementofthebaculovirusexpressionvectorsystembyengineeringanovelexpressioncassette AT josemescribano significantproductivityimprovementofthebaculovirusexpressionvectorsystembyengineeringanovelexpressioncassette |
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