Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light
Chloroplast transformation has many potential advantages for the production of recombinant proteins in plants. However, it has been reported that heterologous protein accumulation in chloroplasts could be hindered by post-transcriptional mechanisms not yet characterized. Here, we describe the develo...
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todo:paper_01681656_v125_n2_p159_Wirth2023-10-03T15:06:05Z Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light Wirth, S. Segretin, M.E. Mentaberry, A. Bravo-Almonacid, F. Chloroplast transformation Fusion protein hEGF Molecular farming Transplastomic tobacco Plants (botany) Tobacco Chloroplast transformation Electron transport inhibitors Fusion protein Human epidermal growth factor (hEGF) Molecular farming Transplastomic tobacco Proteins amino acid bacterial enzyme beta glucuronidase gamma urogastrone hybrid protein protein urea derivative viologen article chloroplast controlled study electron transport genetic transcription incubation time light dark cycle nonhuman nucleotide sequence plant plant leaf priority journal protein expression Blotting, Northern Blotting, Southern Blotting, Western Chloroplasts Epidermal Growth Factor Gene Expression Regulation, Plant Genetic Vectors Glucuronidase Humans Light Plants, Genetically Modified Recombinant Fusion Proteins Tobacco Chlorophylls Plants Plastids Proteins Tobacco Bacteria (microorganisms) Nicotiana obtusifolia Chloroplast transformation has many potential advantages for the production of recombinant proteins in plants. However, it has been reported that heterologous protein accumulation in chloroplasts could be hindered by post-transcriptional mechanisms not yet characterized. Here, we describe the development and characterization of transplastomic tobacco plants transformed with four different transformation vectors for the expression of human epidermal growth factor (hEGF). We showed that, although the corresponding transcript was present in all of the analyzed plants, hEGF could only be detected when fused to the first 186 amino acids of bacterial β-glucuronidase (GUS). In addition, we observed that the expression levels of recombinant protein increased when plants were placed in the dark or when leaves were incubated in the presence of electron transport inhibitors, such as methyl viologen (MV) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). These results suggest that the mechanism responsible for hEGF instability in chloroplasts is regulated by light. © 2006 Elsevier B.V. All rights reserved. Fil:Wirth, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Segretin, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Mentaberry, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_01681656_v125_n2_p159_Wirth |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Chloroplast transformation Fusion protein hEGF Molecular farming Transplastomic tobacco Plants (botany) Tobacco Chloroplast transformation Electron transport inhibitors Fusion protein Human epidermal growth factor (hEGF) Molecular farming Transplastomic tobacco Proteins amino acid bacterial enzyme beta glucuronidase gamma urogastrone hybrid protein protein urea derivative viologen article chloroplast controlled study electron transport genetic transcription incubation time light dark cycle nonhuman nucleotide sequence plant plant leaf priority journal protein expression Blotting, Northern Blotting, Southern Blotting, Western Chloroplasts Epidermal Growth Factor Gene Expression Regulation, Plant Genetic Vectors Glucuronidase Humans Light Plants, Genetically Modified Recombinant Fusion Proteins Tobacco Chlorophylls Plants Plastids Proteins Tobacco Bacteria (microorganisms) Nicotiana obtusifolia |
| spellingShingle |
Chloroplast transformation Fusion protein hEGF Molecular farming Transplastomic tobacco Plants (botany) Tobacco Chloroplast transformation Electron transport inhibitors Fusion protein Human epidermal growth factor (hEGF) Molecular farming Transplastomic tobacco Proteins amino acid bacterial enzyme beta glucuronidase gamma urogastrone hybrid protein protein urea derivative viologen article chloroplast controlled study electron transport genetic transcription incubation time light dark cycle nonhuman nucleotide sequence plant plant leaf priority journal protein expression Blotting, Northern Blotting, Southern Blotting, Western Chloroplasts Epidermal Growth Factor Gene Expression Regulation, Plant Genetic Vectors Glucuronidase Humans Light Plants, Genetically Modified Recombinant Fusion Proteins Tobacco Chlorophylls Plants Plastids Proteins Tobacco Bacteria (microorganisms) Nicotiana obtusifolia Wirth, S. Segretin, M.E. Mentaberry, A. Bravo-Almonacid, F. Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| topic_facet |
Chloroplast transformation Fusion protein hEGF Molecular farming Transplastomic tobacco Plants (botany) Tobacco Chloroplast transformation Electron transport inhibitors Fusion protein Human epidermal growth factor (hEGF) Molecular farming Transplastomic tobacco Proteins amino acid bacterial enzyme beta glucuronidase gamma urogastrone hybrid protein protein urea derivative viologen article chloroplast controlled study electron transport genetic transcription incubation time light dark cycle nonhuman nucleotide sequence plant plant leaf priority journal protein expression Blotting, Northern Blotting, Southern Blotting, Western Chloroplasts Epidermal Growth Factor Gene Expression Regulation, Plant Genetic Vectors Glucuronidase Humans Light Plants, Genetically Modified Recombinant Fusion Proteins Tobacco Chlorophylls Plants Plastids Proteins Tobacco Bacteria (microorganisms) Nicotiana obtusifolia |
| description |
Chloroplast transformation has many potential advantages for the production of recombinant proteins in plants. However, it has been reported that heterologous protein accumulation in chloroplasts could be hindered by post-transcriptional mechanisms not yet characterized. Here, we describe the development and characterization of transplastomic tobacco plants transformed with four different transformation vectors for the expression of human epidermal growth factor (hEGF). We showed that, although the corresponding transcript was present in all of the analyzed plants, hEGF could only be detected when fused to the first 186 amino acids of bacterial β-glucuronidase (GUS). In addition, we observed that the expression levels of recombinant protein increased when plants were placed in the dark or when leaves were incubated in the presence of electron transport inhibitors, such as methyl viologen (MV) and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). These results suggest that the mechanism responsible for hEGF instability in chloroplasts is regulated by light. © 2006 Elsevier B.V. All rights reserved. |
| format |
JOUR |
| author |
Wirth, S. Segretin, M.E. Mentaberry, A. Bravo-Almonacid, F. |
| author_facet |
Wirth, S. Segretin, M.E. Mentaberry, A. Bravo-Almonacid, F. |
| author_sort |
Wirth, S. |
| title |
Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| title_short |
Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| title_full |
Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| title_fullStr |
Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| title_full_unstemmed |
Accumulation of hEGF and hEGF-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| title_sort |
accumulation of hegf and hegf-fusion proteins in chloroplast-transformed tobacco plants is higher in the dark than in the light |
| url |
http://hdl.handle.net/20.500.12110/paper_01681656_v125_n2_p159_Wirth |
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