Expression of two subtypes of human IFN-alpha in transgenic potato plants.
Plant expression systems have advantages over other in vitro expression systems in terms of low production costs and low risk of contamination by animal viruses or bacterial endotoxins. In this study, cDNA encoding two subtypes of human interferon-alpha2b and 8 (HuIFN-alpha2b and HuIFN-alpha8) were introduced into potato plants (Solanum tuberosum) using Agrobacterium-mediated transformation. Transcription and translation of the inserted HuIFN-alpha cDNA were confirmed by Northern blot analysis and ELISA, respectively. Bioactivity of the products was assayed by inhibition of vesicular stomatitis virus (VSV) replication on a human amniotic cell line. However, because of the presence of substances in potato tissue extracts that were toxic to animal cells, successful demonstration of IFN bioactivity in the transformants was achieved only after removal of such substances by dialysis. The maximum level of IFN activity in plant extracts was 560 IU/g of tissue. These results indicated that the HuIFN-alpha gene introduced into the potato plant was correctly translated and transcribed in plant cells. This report for the first time shows that biologically active animal cytokines with potential pharmaceutical applications could be expressed in transgenic potato plants.
Ohya
K
K
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, N18W9, Kita-ku, Sapporo, Hokkaido 060-0818, Japan.
Matsumura
T
T
Ohashi
K
K
Onuma
M
M
Sugimoto
C
C
D000998:Antiviral Agents / Q000494:pharmacology; D015152:Blotting, Northern; D002460:Cell Line; D018076:DNA, Complementary / Q000096:biosynthesis; D018506:Gene Expression Regulation, Plant; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis / Q000494:pharmacology; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000494:pharmacology; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins; D011198:Solanum tuberosum / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection; D014170:Transformation, Genetic
D000998:Antiviral Agents; D018076:DNA, Complementary; D007370:Interferon Type I; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins