@misc{wyatt_tsou_robertson_boss_2004, title={Transgenic plants with increased calcium stores}, volume={6,753,462}, number={2004 June 22}, publisher={Washington, DC: U.S. Patent and Trademark Office}, author={Wyatt, S. and Tsou, P.-L. and Robertson, D. and Boss, W.}, year={2004} } @article{wyatt_tsou_robertson_2002, title={Expression of the high capacity calcium-binding domain of calreticulin increases bioavailable calcium stores in plants}, volume={11}, ISSN={["1573-9368"]}, DOI={10.1023/A:1013917701701}, abstractNote={Modulation of cytosolic calcium levels in both plants and animals is achieved by a system of Ca2+-transport and storage pathways that include Ca2+ buffering proteins in the lumen of intracellular compartments. To date, most research has focused on the role of transporters in regulating cytosolic calcium. We used a reverse genetics approach to modulate calcium stores in the lumen of the endoplasmic reticulum. Our goals were two-fold: to use the low affinity, high capacity Ca2+ binding characteristics of the C-domain of calreticulin to selectively increase Ca2+ storage in the endoplasmic reticulum, and to determine if those alterations affected plant physiological responses to stress. The C-domain of calreticulin is a highly acidic region that binds 20-50 moles of Ca2+ per mole of protein and has been shown to be the major site of Ca2+ storage within the endoplasmic reticulum of plant cells. A 377-bp fragment encoding the C-domain and ER retention signal from the maize calreticulin gene was fused to a gene for the green fluorescent protein and expressed in Arabidopsis under the control of a heat shock promoter. Following induction on normal medium, the C-domain transformants showed delayed loss of chlorophyll after transfer to calcium depleted medium when compared to seedlings transformed with green fluorescent protein alone. Total calcium measurements showed a 9-35% increase for induced C-domain transformants compared to controls. The data suggest that ectopic expression of the calreticulin C-domain increases Ca2+ stores, and that this Ca2+ reserve can be used by the plant in times of stress.}, number={1}, journal={TRANSGENIC RESEARCH}, author={Wyatt, SE and Tsou, PL and Robertson, D}, year={2002}, month={Feb}, pages={1–10} } @article{persson_love_tsou_robertson_thompson_boss_2002, title={When a day makes a difference. Interpreting data from endoplasmic reticulum-targeted green fluorescent protein fusions in cells grown in suspension culture}, volume={128}, ISSN={1532-2548 0032-0889}, url={http://dx.doi.org/10.1104/pp.010840}, DOI={10.1104/pp.010840}, abstractNote={The stability of the self-contained structure of green fluorescent protein (GFP) has made it the most widely utilized fluorescent marker for gene expression and subcellular localization studies ([Chalfie et al., 1994][1]; [Tsien, 1998][2]; [De Giorgi et al., 1999][3]; [Haseloff et al., 1999][4]).}, number={2}, journal={Plant Physiology}, publisher={Oxford University Press (OUP)}, author={Persson, S. and Love, J. and Tsou, P. L. and Robertson, D. and Thompson, W. F. and Boss, W. F.}, year={2002}, pages={341–344} } @article{scott_wyatt_tsou_robertson_allen_1999, title={Model system for plant cell biology: GFP imaging in living onion epidermal cells}, volume={26}, ISSN={["1940-9818"]}, DOI={10.2144/99266st04}, abstractNote={ The ability to visualize organelle localization and dynamics is very useful in studying cellular physiological events. Until recently, this has been accomplished using a variety of staining methods. However, staining can give inaccurate information due to nonspecific staining, diffusion of the stain or through toxic effects. The ability to target green fluorescent protein (GFP) to various organelles allows for specific labeling of organelles in vivo. The disadvantages of GFP thus far have been the time and money involved in developing stable transformants or maintaining cell cultures for transient expression. In this paper, we present a rapid transient expression system using onion epidermal peels. We have localized GFP to various cellular compartments (including the cell wall) to illustrate the utility of this method and to visualize dynamics of these compartments. The onion epidermis has large, living, transparent cells in a monolayer, making them ideal for visualizing GFP. This method is easy and inexpensive, and it allows for testing of new GFP fusion proteins in a living tissue to determine deleterious effects and the ability to express before stable transformants are attempted. }, number={6}, journal={BIOTECHNIQUES}, author={Scott, A and Wyatt, S and Tsou, PL and Robertson, D and Allen, NS}, year={1999}, month={Jun}, pages={1125-+} }