High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.

Tobacco plants can be used to express recombinant proteins that cannot be produced in a soluble and active form using traditional platforms such as Escherichia coli. We therefore expressed the human glycoprotein interleukin 6 (IL6) in two commercial tobacco cultivars (Nicotiana tabacum cv. Virginia...

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Main Authors: Henrik Nausch, Heike Mikschofsky, Roswitha Koslowski, Udo Meyer, Inge Broer, Jana Huckauf
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3495959?pdf=render
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spelling doaj-f58ec409210c4e2480a78d44bdc7ac812020-11-25T01:08:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01711e4893810.1371/journal.pone.0048938High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.Henrik NauschHeike MikschofskyRoswitha KoslowskiUdo MeyerInge BroerJana HuckaufTobacco plants can be used to express recombinant proteins that cannot be produced in a soluble and active form using traditional platforms such as Escherichia coli. We therefore expressed the human glycoprotein interleukin 6 (IL6) in two commercial tobacco cultivars (Nicotiana tabacum cv. Virginia and cv. Geudertheimer) as well as the model host N. benthamiana to compare different transformation strategies (stable vs. transient expression) and subcellular targeting (apoplast, endoplasmic reticulum (ER) and vacuole). In T(0) transgenic plants, the highest expression levels were achieved by ER targeting but the overall yields of IL6 were still low in the leaves (0.005% TSP in the ER, 0.0008% in the vacuole and 0.0005% in the apoplast). The apoplast variant accumulated to similar levels in leaves and seeds, whereas the ER-targeted variant was 1.2-fold more abundant in seeds and the vacuolar variant was 6-fold more abundant in seeds. The yields improved in subsequent generations, with the best-performing T(2) plants producing the ER-targeted IL6 at 0.14% TSP in both leaves and seeds. Transient expression of ER-targeted IL6 in leaves using the MagnICON system resulted in yields of up to 7% TSP in N. benthamiana, but only 1% in N. tabacum cv. Virginia and 0.5% in cv. Geudertheimer. Although the commercial tobacco cultivars produced up to threefold more biomass than N. benthamiana, this was not enough to compensate for the lower overall yields. The recombinant IL6 produced by transient and stable expression in plants was biologically active and presented as two alternative bands matching the corresponding native protein.http://europepmc.org/articles/PMC3495959?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Henrik Nausch
Heike Mikschofsky
Roswitha Koslowski
Udo Meyer
Inge Broer
Jana Huckauf
spellingShingle Henrik Nausch
Heike Mikschofsky
Roswitha Koslowski
Udo Meyer
Inge Broer
Jana Huckauf
High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
PLoS ONE
author_facet Henrik Nausch
Heike Mikschofsky
Roswitha Koslowski
Udo Meyer
Inge Broer
Jana Huckauf
author_sort Henrik Nausch
title High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
title_short High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
title_full High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
title_fullStr High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
title_full_unstemmed High-level transient expression of ER-targeted human interleukin 6 in Nicotiana benthamiana.
title_sort high-level transient expression of er-targeted human interleukin 6 in nicotiana benthamiana.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2012-01-01
description Tobacco plants can be used to express recombinant proteins that cannot be produced in a soluble and active form using traditional platforms such as Escherichia coli. We therefore expressed the human glycoprotein interleukin 6 (IL6) in two commercial tobacco cultivars (Nicotiana tabacum cv. Virginia and cv. Geudertheimer) as well as the model host N. benthamiana to compare different transformation strategies (stable vs. transient expression) and subcellular targeting (apoplast, endoplasmic reticulum (ER) and vacuole). In T(0) transgenic plants, the highest expression levels were achieved by ER targeting but the overall yields of IL6 were still low in the leaves (0.005% TSP in the ER, 0.0008% in the vacuole and 0.0005% in the apoplast). The apoplast variant accumulated to similar levels in leaves and seeds, whereas the ER-targeted variant was 1.2-fold more abundant in seeds and the vacuolar variant was 6-fold more abundant in seeds. The yields improved in subsequent generations, with the best-performing T(2) plants producing the ER-targeted IL6 at 0.14% TSP in both leaves and seeds. Transient expression of ER-targeted IL6 in leaves using the MagnICON system resulted in yields of up to 7% TSP in N. benthamiana, but only 1% in N. tabacum cv. Virginia and 0.5% in cv. Geudertheimer. Although the commercial tobacco cultivars produced up to threefold more biomass than N. benthamiana, this was not enough to compensate for the lower overall yields. The recombinant IL6 produced by transient and stable expression in plants was biologically active and presented as two alternative bands matching the corresponding native protein.
url http://europepmc.org/articles/PMC3495959?pdf=render
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