Rapid transient expression of human granulocyte-macrophage colony-stimulating factor in two industrial cultivars of tobacco (Nicotiana tabacum L.) by agroinfiltration

• Successful expression of human cytokine GM-CSF in two industrial tobacco cultivars is achieved.
• Rapid cloning in two binary destination vectors is accomplished by using Gateway approach.
• Agrobacterial infiltration procedure is optimized and shown to require the surface tension lowering agent Silwet L-77.
• Accumulation of recombinant protein was confirmed by using high affinity monoclonal hemagglutinin tag antibodies.
• Production of hGM-CSF has been achieved without plant codon usage optimization.

We report the production of hGM-CSF cytokine in leaves of industrial tobacco cultivars DH-17 and DH-27 by using Agrobacterium-mediated transient expression. We prove the concept that very high biomass industrial tobacco plants are suitable platforms for rapid, low cost production of foreign proteins. Successful transient expression of the GM-CSF was achieved in less than three months, opening the possibility for future applications of this approach in rapid response production of various proteins of non-plant origin in industrial tobacco.