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

(2002) NATURE BIOTECHNOLOGY. 20(12). p.1265-1268
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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.
Keywords
PETUNIA-HYBRIDA, SCFV ANTIBODY, CYTOSOL, AGROBACTERIUM-MEDIATED TRANSFORMATION, ACUTIFOLIUS, CHAIN VARIABLE FRAGMENTS, GENE, EXPRESSION, PLANTS, LEVEL

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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.
@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},
}

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