Molecular farming of human tissue transglutaminase in tobacco plants

In this study we have utilized Nicotiana tabacum with a molecular farming purpose in attempt of producing transgenic plants expressing the human tissue transglutaminase (htTG). Three plant expression constructs were used enabling targeting and accumulation of the recombinant protein into the plant cell cytosol (cyto), the chloroplasts (chl) and the apoplastic space (apo). Analysis of transgenic T-0 plants revealed that recombinant htTG was detectable in all three transgenic lines and the accumulation levels were in a range of 18-75 mu g/g of leaf material. In the T-1 generation, the recombinant htTG was still expressed at high level and a significant catalytic activity was detected into the leaf protein extracts. Southern blot analyses revealed that apo and chl plants of T-1 generation possess a high copy number of the recombinant htTG in their genome, while the cyto plants carry a single copy.