Sorting of Glycoprotein B from Human Cytomegalovirus to Protein Storage Vesicles in Seeds of Transgenic Tobacco |
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Authors: | Wright KE Prior F Sardana R Altosaar I Dudani AK Ganz PR Tackaberry ES |
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Institution: | (1) Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Canada;(2) Bureau of Biologics and Radiopharmaceuticals, Therapeutic Products Programme, Health Canada, Tunney s Pasture 2201C, Ottawa, Ontario, Canada, K1A 0L2;(3) Department of Cellular and Molecular Medicine, University of Ottawa, Canada |
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Abstract: | As part of ongoing studies into the use of plant expression systems for making human therapeutic proteins, we have successfully expressed the major glycoprotein, gB, of human cytomegalovirus (HCMV) in transgenic tobacco plants. Viral glycoprotein was detectable in the protein extracts of mature tobacco seeds using neutralizing and non-neutralizing monoclonal antibodies specific for gB. Although several mammalian proteins have been expressed in tobacco, localization of these proteins in transgenic tobacco tissue has not been extensively examined. The objective of this study was to identify the site(s) of recombinant gB deposition in mature tobacco seeds. Using immunogold labelling and electron microscopy, we found specific labelling for gB in the endosperm of transgenic seeds, with gB localized almost exclusively in protein storage vesicles (PSV). This occurred in seeds that were freshly harvested and in seeds that had been stored for several months. These data indicate that gB behaves like a plant storage protein when expressed in tobacco seeds, and provide further support for the suitability of plants for producing recombinant proteins of potential clinical relevance. |
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Keywords: | viral glycoprotein human cytomegalovirus transgenic tobacco seeds protein storage vesicles |
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