Recombinant mouse granulocyte-macrophage colony-stimulating factor is glycosylated in transgenic tobacco and maintains its biological activity
| Autor: | Agnieszka Sirko, Dali Gaganidze, Robert Brodzik, Anna Góra-Sochacka, Patrycja Redkiewicz, Bogusława Napiórkowska |
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| Rok vydání: | 2009 |
| Předmět: |
Macrophage colony-stimulating factor
Glycosylation Transgene Nicotiana tabacum KDEL Immunology Blotting Western Biology law.invention Cell Line Mice law Virology Complementary DNA Tobacco Animals Endoplasmic reticulum fungi food and beverages Granulocyte-Macrophage Colony-Stimulating Factor Cell Biology biology.organism_classification Blotting Northern Plants Genetically Modified Molecular biology Recombinant Proteins Cell culture Recombinant DNA Plasmids |
| Zdroj: | Journal of interferoncytokine research : the official journal of the International Society for Interferon and Cytokine Research. 30(3) |
| ISSN: | 1557-7465 |
| Popis: | The granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine with many important applications and, due to its immunostimulatory properties, could also be used as a vaccine adjuvant. A simple strategy to produce recombinant mouse GM-CSF (mGM-CSF) in transgenic Nicotiana tabacum plants was used in this study. The mGM-CSF cDNA followed by the sequence encoding endoplasmic reticulum retention signal (KDEL) was cloned into the ImpactVector under the control of the strong promoter from the gene encoding a small subunit of Rubisco. In transgenic plants the accumulation level of recombinant mGM-CSF varied in the individual transformants from 8 to 19 microg/g of fresh leaf tissue, which makes up to 0.22% of total soluble protein. In most analyzed plants, the apparent molecular weight of the recombinant protein was larger than predicted due to its N-glycosylation, presumably in 2 sites. The recombinant plant-produced murine GM-CSF retained its biological activity as confirmed in vitro in proliferation assay using a mouse cell line, which is growth-dependent on GM-CSF. |
| Databáze: | OpenAIRE |
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