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Vacuole accumulation of storage protein and lectin expressed in transgenic tobacco seeds
Institution:1. Plant Molecular Biology Laboratory, United States Department of Agriculture, Agricultural Research Service, Beltsville, MD 20705, U.S.A.;2. Biology Department C-016, University of California, San Diego, CA, U.S.A. 92093;3. Agrigenetics Advanced Science Company, Madison, WI 53716, U.S.A.;1. Department of Agroecology, Faculty of Science and Technology, Aarhus University, P.O. Box 50, DK-8830 Tjele, Denmark;2. Department of Animal Science, Faculty of Science and Technology, Aarhus University, Denmark;3. Danish Centre for Food and Agriculture, Aarhus University, Denmark;1. Institute of Animal Nutrition and Functional Plant Compounds, Department for Farm Animals and Veterinary Public Health, Vetmeduni Vienna, Veterinaerplatz 1, 1210 Vienna, Austria;2. Institute of Animal Nutrition, Livestock Products and Nutrition Physiology, University of Natural Resources and Life Sciences Vienna, Department for Agrobiotechnology, Muthgasse 11, 1190 Vienna, Austria;1. Animal & Grassland Research and Innovation Centre, Teagasc Moorepark, Cork, Ireland;2. LUKE Agrifood Research Finland, FI-31600 Jokioinen, Finland;3. Department of Agricultural Science, University of Helsinki, Finland
Abstract:The expression of proteins in transgenic plants offers an elegant means to examine targeting signals used for transport to intracellular sites of accumulation. We have used electron microscopic immunogold procedures to localize several different storage proteins and lectins expressed in transgenic tobacco seeds. The objective of these studies is to characterize targeting signals which permit translocation and accumulation in protein storage vacuoles (protein bodies). Vacuolar proteins such as phaseolin and phytohemagglutinin (PHA) are correctly transported to the protein storage vacuoles of transgenic tobacco seeds. Site-directed mutagenesis was used to change Asn-linked glycosylation sites of PHA. Minus glycan PHA was accumulated in the protein storage vacuoles indicating that glycans do not confer targeting information. Zein the non-vacuolar storage protein of maize accumulates in the protein storage vacuoles indicating that deposition occurs in some proteins which may lack vacuolar targeting signals.
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