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Boosting heterologous protein production in transgenic dicotyledonous seeds using Phaseolus vulgaris regulatory sequences

Geert De Jaeger UGent, Stanley Scheffer, Anni Jacobs UGent, Mukund Zambre UGent, Oliver Zobell, Alain Goossens UGent, Anna Depicker UGent and Geert Angenon UGent (2002) NATURE BIOTECHNOLOGY. 20(12). p.1265-1268
abstract
Over the past decade, several high-value proteins have been produced in different transgenic plant tissues such as leaves, tubers, and seeds(1,2). Despite recent advances, many heterologous proteins accumulate to low concentrations, and the optimization of expression cassettes to make in planta production and purification economically feasible remains critical. Here, the regulatory sequences of the seed storage protein gene arcelin 5-I (arc5-I) of common bean (Phaseolus vulgaris)(3) were evaluated for producing heterologous proteins in dicotyledonous seeds. The murine single-chain variable fragment (scFv) G4 (ref. 4) was chosen as model protein because of the current industrial interest in producing antibodies and derived fragments in crops(5,6). In transgenic Arabidopsis thaliana seed stocks, the scFv under control of the 35S promoter of the cauliflower mosaic virus (CaMV) accumulated to approximately 1% of total soluble protein (TSP). However, a set of seed storage promoter constructs boosted the scFv accumulation to exceptionally high concentrations, reaching no less than 36.5% of TSP in homozygous seeds. Even at these high concentrations, the scFv proteins had antigen-binding activity and affinity similar to those produced in Escherichia coli. The feasibility of heterologous protein production under control of arc5-I regulatory sequences was also demonstrated in Phaseolus acutifolius, a promising crop for large-scale production.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
PETUNIA-HYBRIDA, SCFV ANTIBODY, CYTOSOL, AGROBACTERIUM-MEDIATED TRANSFORMATION, ACUTIFOLIUS, CHAIN VARIABLE FRAGMENTS, GENE, EXPRESSION, PLANTS, LEVEL
journal title
NATURE BIOTECHNOLOGY
Nat. Biotechnol.
volume
20
issue
12
pages
1265 - 1268
Web of Science type
Article
Web of Science id
000179552600049
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
12.822 (2002)
JCR rank
1/130 (2002)
JCR quartile
1 (2002)
ISSN
1087-0156
DOI
10.1038/nbt755
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
153867
handle
http://hdl.handle.net/1854/LU-153867
date created
2004-01-14 13:38:00
date last changed
2017-03-02 15:48:08
@article{153867,
  abstract     = {Over the past decade, several high-value proteins have been produced in different transgenic plant tissues such as leaves, tubers, and seeds(1,2). Despite recent advances, many heterologous proteins accumulate to low concentrations, and the optimization of expression cassettes to make in planta production and purification economically feasible remains critical. Here, the regulatory sequences of the seed storage protein gene arcelin 5-I (arc5-I) of common bean (Phaseolus vulgaris)(3) were evaluated for producing heterologous proteins in dicotyledonous seeds. The murine single-chain variable fragment (scFv) G4 (ref. 4) was chosen as model protein because of the current industrial interest in producing antibodies and derived fragments in crops(5,6). In transgenic Arabidopsis thaliana seed stocks, the scFv under control of the 35S promoter of the cauliflower mosaic virus (CaMV) accumulated to approximately 1\% of total soluble protein (TSP). However, a set of seed storage promoter constructs boosted the scFv accumulation to exceptionally high concentrations, reaching no less than 36.5\% of TSP in homozygous seeds. Even at these high concentrations, the scFv proteins had antigen-binding activity and affinity similar to those produced in Escherichia coli. The feasibility of heterologous protein production under control of arc5-I regulatory sequences was also demonstrated in Phaseolus acutifolius, a promising crop for large-scale production.},
  author       = {De Jaeger, Geert and Scheffer, Stanley and Jacobs, Anni and Zambre, Mukund and Zobell, Oliver and Goossens, Alain and Depicker, Anna and Angenon, Geert},
  issn         = {1087-0156},
  journal      = {NATURE BIOTECHNOLOGY},
  keyword      = {PETUNIA-HYBRIDA,SCFV ANTIBODY,CYTOSOL,AGROBACTERIUM-MEDIATED TRANSFORMATION,ACUTIFOLIUS,CHAIN VARIABLE FRAGMENTS,GENE,EXPRESSION,PLANTS,LEVEL},
  language     = {eng},
  number       = {12},
  pages        = {1265--1268},
  title        = {Boosting heterologous protein production in transgenic dicotyledonous seeds using Phaseolus vulgaris regulatory sequences},
  url          = {http://dx.doi.org/10.1038/nbt755},
  volume       = {20},
  year         = {2002},
}

Chicago
De Jaeger, Geert, Stanley Scheffer, Anni Jacobs, Mukund Zambre, Oliver Zobell, Alain Goossens, Anna Depicker, and Geert Angenon. 2002. “Boosting Heterologous Protein Production in Transgenic Dicotyledonous Seeds Using Phaseolus Vulgaris Regulatory Sequences.” Nature Biotechnology 20 (12): 1265–1268.
APA
De Jaeger, G., Scheffer, S., Jacobs, A., Zambre, M., Zobell, O., Goossens, A., Depicker, A., et al. (2002). Boosting heterologous protein production in transgenic dicotyledonous seeds using Phaseolus vulgaris regulatory sequences. NATURE BIOTECHNOLOGY, 20(12), 1265–1268.
Vancouver
1.
De Jaeger G, Scheffer S, Jacobs A, Zambre M, Zobell O, Goossens A, et al. Boosting heterologous protein production in transgenic dicotyledonous seeds using Phaseolus vulgaris regulatory sequences. NATURE BIOTECHNOLOGY. 2002;20(12):1265–8.
MLA
De Jaeger, Geert, Stanley Scheffer, Anni Jacobs, et al. “Boosting Heterologous Protein Production in Transgenic Dicotyledonous Seeds Using Phaseolus Vulgaris Regulatory Sequences.” NATURE BIOTECHNOLOGY 20.12 (2002): 1265–1268. Print.