Gora-Sochacka A.,Polish Academy of Sciences |
Redkiewicz P.,Polish Academy of Sciences |
Napiorkowska B.,Polish Academy of Sciences |
Napiorkowska B.,Research Institute of Pomology and Floriculture |
And 4 more authors.
Journal of Interferon and Cytokine Research | Year: 2010
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 g/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.© Mary Ann Liebert, Inc. 2009.