Improved Production of the Human Granulocyte-Macrophage Colony Stimulating Factor in Transgenic Arabidopsis thaliana Seeds Using a Dual Sorting Signal Peptide

Abstract

The human granulocyte-macrophage colony stimulating factor (hGM-CSF) containing either an endoplasmic reticulum (ER) retention signal or a phaseolin vacuolar sorting signal peptide was expressed in Arabidopsis thaliana under the control of a tissue-specific promoter, derived from the soybean α′ subunit of β-conglycinin. No significant differences in recombinant hGM-CSF (rhGM-CSF) accumulation were detected between transgenic plants carrying either one of the two signal peptides. Hybrid seed from crosses between single-copy transformants tailed with the ER retention signal tetrapeptide and single-copy transformed plants tagged with a phaseolin four carboxy-terminal residues showed gene additive effects. The highest expression level of rhGM-CSF was 0.05% of total soluble protein of immature siliques, indicating that the two signal peptides functioned independently in the protein-sorting pathway. Additionally, TF-1 cell proliferation data demonstrated that rhGM-CSF was biologically active.

Transgenic Soybean for Production of Thermostable α-Amylase

“…The AmyS expression cassette is composed of a seed-specific promoter, the coding sequence for the Arabidopsis 2S albumin (At2S) signal peptide and AmyS, and the Arabidopsis heat shock protein 18.2 (AtHSP) terminator. The At2S signal peptide is anticipated to direct the synthesized protein to the endoplasmic reticulum [ 21 , 22 , 23 ]. The Nicotiana tabacum Rb7 matrix attachment region (MAR) was added to the 3’ end of the AmyS expression cassette.…”

Expression of the Human Tissue-Plasminogen Activator in Hairy Roots of Oriental Melon (Cucumis melo)

“…Many therapeutic proteins, valuable secondary metabolites and functional recombinant proteins have been produced using hairy roots cultures or transgenic plants, such as thymosin alpha 1 (Cui et al 2010), human secreted alkaline phosphatase (Gaume et al 2003), fibrinolytic enzyme ), elevated ginsenoside (Liang et al 2009), cry1Ac (Nandeshwar et al 2009), monoclonal antibodies (Sharp and Doran 2001), synthetic heat-labile enterotoxin B subunit and hemagglutinin-neuraminidaseneutralizing epitope fusion protein ), ribosome-inactivating proteins (Thorup et al 1994), human acetylcholinesterase (Woods et al 2008), hepatitis B surface antigen (Unni and Soniya 2010), and human granulocyte-macrophage colony stimulating factor (Wang et al 2009). These studies demonstrate that hairy roots and transgenic plants can be a better alternative to mammalian cell cultures and microbial fermentation for the production of recombinant proteins.…”