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Free Content Virus-mediated delivery of the green fluorescent protein to the endoplasmic reticulum of plant cells

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The green fluorescent protein (GFP) was targeted to the endoplasmic reticulum (ER) of living plant cells using a virus-based expression system. The signal peptide from the storage protein, patatin was fused to the amino-terminus of the GFP while the ER retention signal KDEL was fused to the carboxy-terminus. The chimaeric gfp cDNA was inserted into a potato virus X-based expression vector and in-vitro transcripts, representing the recombinant viral genome, were inoculated on to Nicotiana benthamiana and N. clevelandii plants. In virus-infected cells, the GFP was targeted both to the cortical ER and to a mobile system of ER elements which underwent streaming in the cytoplasm. In addition, a population of GFP-containing inclusions was apparent. These inclusions were motile but remained closely associated with elements of the ER. Staining of the ER with membrane potential-sensitive dyes confirmed that the GFP had been targeted to the ER. The utility of virus-mediated delivery systems in studies of the plant endomembrane system is discussed.
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Document Type: Research Article

Affiliations: 1: Unit of Plant Transport Processes, Department of Cellular and Environmental Physiology, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK 2: School of Biological and Molecular Sciences, Oxford Brookes University, Gypsy Lane, Oxford OX3 0BP, UK 3: Department of Biological Sciences, University of Durham, South Road, Durham DH1 3LE, UK

Publication date: November 1, 1996

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