Live imaging and sub-cellular localization of the a9-gus-gfp chimeric protein expressed in nicotiana benthamiana transgenic plants

Abstract

The fusion of the signal peptide region from the Arabidopsis A9 anther-specific gene to a GUS-GFP reporter protein results in direction of the chimeric protein to ER and its further secretion out of the cell, following expression in Nicotiana benthamiana cell suspension. The detailed observations of the sub-cellular translocation and exocytosis of the chimeric A9-GUS-GFP protein have been largely restricted by the strong fluorescence originating from to the accumulation of high levels of the fluorescent protein in the endo-membrane compartments and plasma membrane. We report using of cold-pretreatment and photo bleaching for more detail fluorescent microscopy observation of the sub-cellular compartmentalization and secretion pattern of the A9-GUS-GFP protein in leaf cells of the transgenic N. benthamiana plants. © 2008 Taylor and Francis Group, LLC.