Identification of the factors that control synthesis and accumulation of a therapeutic protein, human immune-regulatory interleukin-10, in Arabidopsis thaliana
Ling Chen
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorBrian R. Dempsey
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorLaszlo Gyenis
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorRima Menassa
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorJim E. Brandle
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorCorresponding Author
Sangeeta Dhaubhadel
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Correspondence (Tel 519 457 1470 ext. 670; fax 519 457 3997; email [email protected])Search for more papers by this authorLing Chen
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorBrian R. Dempsey
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorLaszlo Gyenis
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorRima Menassa
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorJim E. Brandle
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Search for more papers by this authorCorresponding Author
Sangeeta Dhaubhadel
Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada
Correspondence (Tel 519 457 1470 ext. 670; fax 519 457 3997; email [email protected])Search for more papers by this authorSummary
Plants are one of the most economical platforms for large-scale production of recombinant proteins for biopharmaceutical and industrial uses. A large number of human recombinant proteins of therapeutic value have been successfully produced in plant systems. One of the main technical challenges of producing recombinant proteins in plants is to obtain sufficient level of protein. This research aims to identify the factors that control synthesis and accumulation of recombinant proteins in stable transgenic plants. A stepwise dissection of human immune-regulatory interleukin-10 (IL-10) protein production was carried out using Arabidopsis thaliana as a model system. EMS-mutagenized transgenic Arabidopsis IL-10 lines, at2762 and at3262, produced significantly higher amount of IL-10 protein than the non-mutagenized IL-10 line (WT-IL-10). The fates of trans-gene in these sets of plants were compared in detail by measuring synthesis and accumulation of IL-10 transcript, transcript stability, protein synthesis and IL-10 protein accumulation. The IL-10 transcripts were more stable in at2762 and at3262 lines than WT-IL-10, which may contribute to higher protein synthesis in these lines. To evaluate whether translational regulation of IL-10 controls its synthesis in non-mutagenized WT-IL-10 and higher IL-10 accumulating mutant lines, we measured the efficiency of the translational machinery. Our results indicate that mutant lines with higher trans-gene expression contain more robust and efficient translational machinery compared with the control line.
Supporting Information
| Filename | Description |
|---|---|
| pbi12042-sup-0001-FigS1.pdfapplication/PDF, 945.2 KB | Figure S1 Kinetics of IL-10 mRNA decay in WT-IL-10 and mutant lines at3262 and at2762. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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