Expression of biologically active human tumor necrosis factor-alpha in transgenic potato plant.
We report the successful insertion of the cDNA of human tumor necrosis factor-alpha (HuTNF-alpha) into the genome of potato plant species, Solanum tuberosum, using Agrobacterium tumefacience-mediated transformation. HuTNF-alpha is a known and essential cytokine mediating host defense against tumors and infectious diseases and an immunomodulating agent. To enhance the accumulation of foreign gene product expression in plant cells, the molecular design of the constructed HuTNF-alpha is presented. Transcription and translation of TNF-alpha in transformants were confirmed by Northern blot, RT-PCR, ELISA, and Western blot, respectively. Expression of the bioactive HuTNF-alpha in plant cells was confirmed by way of the cytotoxic effect of the extract obtained from the transformants against murine L929 cells. We think that the expression level of HuTNF-alpha (15 microg/g potato plant tissue) obtained in the present study may be sufficient to induce responses/effects similar to those generated by TNF-alpha in human milk administered orally. We believe that the TNF-alpha expressed in edible potato plants has tremendous potential for clinical use in the areas of medicine and veterinary science. Its usefulness and applicability, therefore, need to be fully explored.
Ohya
Kenji
K
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku, Sapporo 060-0818, Hokkaido, Japan.
Itchoda
Noriko
N
Ohashi
Kazuhiko
K
Onuma
Misao
M
Sugimoto
Chihiro
C
Matsumura
Takeshi
T
D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D019677:Artificial Gene Fusion; D018506:Gene Expression Regulation, Plant; D018745:Genome, Plant; D006801:Humans; D007739:L Cells; D051379:Mice; D030821:Plants, Genetically Modified; D011198:Solanum tuberosum / Q000235:genetics* / Q000378:metabolism*; D014170:Transformation, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
D014409:Tumor Necrosis Factor-alpha