Tobacco as biofactory for biologically active hPL production: a human hormone with potential applications in type-1 diabetes.
Human placental lactogen (hPL) is a peptidic hormone that belongs to the short list of growth factors that could treat type-1 diabetes through pancreatic islet transplantation. Placental lactogen has the capacity to improve islet survival and function before or after transplantation. In this study, transgenic tobacco plants were used as a novel expression system for the production of recombinant hPL protein (rhPL). The expression vector pNEKhPL2 containing hPL cDNA was introduced into tobacco plants; the transcriptional activity was confirmed by real-time PCR, and the rhPL levels reached 1% of the total soluble protein (TSP) content in plants cultivated in the greenhouse. In vitro bioassays using the rat insulinoma (INS-1) cell line showed that recombinant protein was able to induce cell proliferation and activate the JAK-2/STAT-5 signal transduction pathway, demonstrating that plant cells can produce the biologically active hPL protein. To further characterize the plant expression system for hPL production, we analyzed the stability of the protein during the life cycle of tobacco plants as well as the transmission of the transgenic trait to the progeny. The recombinant protein was stably accumulated in young leaves, reaching the maximum level in the first month (6.51 μg/g of fresh weight), but showing a decreasing trend of 26% from the initial sampling time until the end of plant's life cycle. The progeny of the selected pNEKhPL2 plant showed in vitro expression levels of up to 1.1% of TSP. Our results therefore indicate that transgenic plants are a suitable expression system for hPL production.
Neiker-Tecnalia, Arkaute, PO Box 46, E-01080, Vitoria-Gasteiz, Spain.
Iratxe
I
Urreta
Iñigo
I
Oyanguren
Sonia
S
Castañón
D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003922:Diabetes Mellitus, Type 1 / Q000188:drug therapy / Q000601:surgery; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D050417:Insulin-Secreting Cells / Q000187:drug effects*; D016381:Islets of Langerhans Transplantation; D010928:Placental Lactogen / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D030821:Plants, Genetically Modified; D055550:Protein Stability; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D015398:Signal Transduction; D014026:Nicotiana / Q000235:genetics*
D011994:Recombinant Proteins; D010928:Placental Lactogen