Miao Yansong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 78-83). / Abstracts in English and Chinese. / Thesis/Assessment Committee --- p.ii / Statement --- p.iii / 摘要 --- p.vi / Acknowledgements --- p.vii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1. --- The plant secretory pathway --- p.2 / Chapter 2. --- Two different types of vacuoles in plant cells --- p.2 / Chapter 3. --- Vacuolar sorting receptor (VSR) proteins --- p.3 / Chapter 4. --- BP-80 and prevacuolar compartment --- p.6 / Chapter 5. --- The Arabidopsis VSR proteins --- p.7 / Chapter 6. --- Research objectives --- p.8 / Chapter Chapter 2 --- Development of Transgenic Tobacco BY-2 Cell Lines Expressing GFP-AtVSR Fusions --- p.10 / Chapter 1. --- Introduction --- p.11 / Chapter 2. --- Materials and Methods --- p.12 / Chapter 2.1 --- Structure of Golgi marker and PVC marker --- p.12 / Chapter 2.2 --- Construction of GFP-VSR reporters --- p.14 / Chapter 2.3 --- Agrobacterium electroporation --- p.24 / Chapter 2.4 --- Transformation of tobacco BY-2 cells --- p.24 / Chapter 2.5 --- Screening of transgenic BY-2 cells expressing GFP-VSR fusions --- p.25 / Chapter 2.6 --- Chemicals --- p.27 / Chapter 3. --- Result.s --- p.28 / Chapter 3.1 --- Chimeric GFP reporters as tools to study subcellular localization of Arabidopsis vacuolar sorting receptor proteins in transgenic BY-2 cells --- p.28 / Chapter 3.2 --- Establishment of transgenic tobacco (Nicotiana tabacum) BY-2 cell lines stably expressing seven GFP-AtVSR reporters --- p.29 / Chapter 4. --- Conclusion --- p.38 / Chapter Chapter 3 --- Subcellular Localization of the Seven GFP-AtVSR Fusions to Prevacuolar Compartments in Transgenic Tobacco BY-2 Cells --- p.39 / Chapter 1. --- Introduction --- p.40 / Chapter 2. --- Materials and Methods --- p.41 / Chapter 2.1 --- Confocal immunofluorescence studies --- p.41 / Chapter 2.2 --- Antibodies for immunolabeling --- p.42 / Chapter 2.3 --- Wortmannin and brefeldin A treatment --- p.42 / Chapter 2.4 --- Electron microscopy of resin-embedded cells --- p.43 / Chapter 3. --- Results --- p.44 / Chapter 3.1 --- Vacuolar sorting receptor proteins in plants --- p.44 / Chapter 3.2 --- PVC localization of GFP-AtVSR fusions in transgenic tobacco BY-2 cells --- p.47 / Chapter 3.3 --- The spacer sequences did not affect PVC localization of GFP-AtVSR7 --- p.56 / Chapter 3.4 --- Wortmannin-induced vacuolated PVCs contained VSRs in tobacco BY-2 cells --- p.58 / Chapter 3.5 --- Wortmannin-induced vacuolation of PVCs is a general response in plant cells --- p.62 / Chapter 4. --- Conclusion --- p.65 / Chapter Chapter 4 --- Discussion and Future Perspectives --- p.66 / Chapter 1. --- The hypothesis in this study --- p.67 / Chapter 2. --- GFP and BY-2 cells --- p.67 / Chapter 3. --- A reporter system to study subcellular localization of VSR proteins in transgenic tobacco BY-2 cells --- p.68 / Chapter 4. --- PVC localization of the seven GFP-AtVSR fusions in transgenic BY-2 cells --- p.69 / Chapter 5. --- VSR spacer sequences did not affect PVC localization of GFP-AtVSR fusions in transgenic tobacco BY-2 cells --- p.71 / Chapter 6. --- PVC localization of GFP-PV72 and GFP-AtVSR 1 fusions in transgenic tobacco BY-2 cells --- p.73 / Chapter 7. --- Wortmannin-induced vacuolation of PVC is a general response in plant cells --- p.75 / Chapter 8. --- Future perspectives --- p.75 / References --- p.78