Fung Ka Leung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 83-93). / Abstracts in English and Chinese. / Thesis Committee --- p.ii / Statement --- p.iii / Acknowledgements --- p.iv / Abstract --- p.v / 摘要 --- p.vii / Table of Contents --- p.viii / List of Tables --- p.xii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- An overview of the secretory pathway in eukaryotic cells --- p.2 / Chapter 1.2 --- The secretory pathway in plants --- p.4 / Chapter 1.2.1 --- Plant cells contain two functionally distinct vacuoles --- p.4 / Chapter 1.2.2 --- Three vesicular pathways to two vacuole --- p.6 / Chapter 1.2.3 --- Transport vesicles in the three vesicular pathways --- p.9 / Chapter 1.2.4 --- Vacuolar sorting determinants (VSDs) --- p.10 / Chapter 1.2.5 --- Vacuolar sorting receptors (VSRs) --- p.12 / Chapter 1.3 --- The PSVs in mature seeds --- p.15 / Chapter 1.3.1 --- Biogenesis of PSV --- p.15 / Chapter 1.3.2 --- The two chimeric integral membrane reporters --- p.16 / Chapter 1.3.3 --- Subcellular localization of the two chimeric integral membrane reporters in PSVs of mature tobacco seeds --- p.17 / Chapter 1.4 --- Project objectives --- p.19 / Chapter Chapter 2 --- Materials and Methods --- p.20 / Chapter 2.1 --- Construction of the YFP-BP-80 and the YFP- a -TIP reporters --- p.21 / Chapter 2.1.1 --- The pYFP-BP-80-K construct --- p.21 / Chapter 2.1.2 --- The pYFP- a -TIP-K construct --- p.22 / Chapter 2.2 --- Construction of GFP-RMR reporter --- p.23 / Chapter 2.2.1 --- Cloning of pGFP-RMR --- p.23 / Chapter 2.2.2 --- Cloning of pGFP-RMR-K --- p.23 / Chapter 2.3 --- Construction of pGONST1-YFP construct --- p.26 / Chapter 2.3.1 --- The pGONSTl-YFP construct --- p.26 / Chapter 2.4 --- Transformation of Agrobacterium by electroporation --- p.27 / Chapter 2.5 --- Tobacco transformation and selection --- p.28 / Chapter 2.5.1 --- Plant materials --- p.28 / Chapter 2.5.2 --- Tobacco transformation --- p.28 / Chapter 2.6 --- Screening of transgenic tobacco plants expressing YFP fusion proteins --- p.30 / Chapter 2.6.1 --- Kanamycin screening --- p.30 / Chapter 2.6.2 --- Extraction of genomic DNA from leaves --- p.30 / Chapter 2.6.3 --- PCR of genomic DNA --- p.31 / Chapter 2.7 --- Southern blot analysis of genomic DNA --- p.32 / Chapter 2.8 --- Western blot analysis of transgenic tobacco plants --- p.33 / Chapter 2.8.1 --- Extraction of total protein from tobacco leaves or seeds --- p.33 / Chapter 2.8.2 --- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis --- p.34 / Chapter 2.9 --- Confocal immunofluorescence studies --- p.35 / Chapter 2.9.1 --- Preparation of sections --- p.35 / Chapter 2.9.2 --- Single labeling --- p.35 / Chapter 2.9.3 --- Double labeling with one polyclonal and one monoclonal antibodies --- p.36 / Chapter 2.9.4 --- Double labeling with two polyclonal antibodies --- p.36 / Chapter 2.9.5 --- Collection of images --- p.37 / Chapter 2.10 --- Chemicals --- p.38 / Chapter 2.11 --- Primers --- p.38 / Chapter 2.12 --- Bacterial strain --- p.38 / Chapter 2.13 --- Antibodies --- p.39 / Chapter 2.14 --- Growing condition of transgenic plants and determining the developmental stage of tobacco flowers --- p.39 / Chapter Chapter 3 --- Results --- p.41 / Chapter 3.1 --- Generation of transgenic tobacco plants --- p.42 / Chapter 3.2 --- PCR screening of transgenic tobacco plants --- p.46 / Chapter 3.3 --- Southern blot analysis --- p.48 / Chapter 3.4 --- Detection of the YFP fusion proteins in transgenic tobacco plants by western blot analysis --- p.50 / Chapter 3.4.1 --- Detection of the YFP fusion proteins in leaves --- p.50 / Chapter 3.4.2 --- Western blot analysis of vegetative tissues --- p.57 / Chapter 3.4.3 --- Western blot analysis of mature seeds --- p.59 / Chapter 3.5 --- Confocal immunofluorescence studies --- p.61 / Chapter 3.5.1 --- Detection of YFP signals in root tip cells --- p.61 / Chapter 3.5.2 --- Detection of YFP signals in developing seeds --- p.65 / Chapter 3.5.3 --- Subcellular localization of the YFP fusion proteins in mature seeds --- p.67 / Chapter Chapter 4 --- Discussion --- p.72 / Chapter Chapter 5 --- Summary and Future Perspectives --- p.77 / Chapter 5.1 --- Summary --- p.78 / Chapter 5.1.1 --- Generation of transgenic tobacco plants expressing the YFP fusion proteins --- p.78 / Chapter 5.1.2 --- Full-length fusion proteins and cleaved soluble YFP were detected in vegetative tissues --- p.79 / Chapter 5.1.3 --- Only cleaved soluble YFP was detected in mature seeds --- p.79 / Chapter 5.1.4 --- The two fusion proteins might localized in different compartments in developing seeds --- p.79 / Chapter 5.1.5 --- Both fusion proteins were localized within the PSVs of mature seeds --- p.80 / Chapter 5.2 --- Future perspectives --- p.81 / References --- p.83

by Tsang Kwok Yeung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 82-93). / ACKNOWLEDGMENTS --- p.I / ABSTRACT --- p.II / TABLE OF CONTENTS --- p.V / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- PURIFICATION AND CHARACTERIZATION OF A TYPE I RIP MOMORGROSVIN FROM THE SEEDS OF MOMORDICA GROSVENORI --- p.34 / GENERAL DISCUSSION --- p.80 / REFERENCES --- p.82

by Tse Man Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 74-81). / ACKNOWLEDGMENTS --- p.I / ABSTRACT --- p.II / LIST OF ABBREVIATIONS --- p.III / TABLE OF CONTENTS --- p.1 / Chapter CHAPTER 1 --- INTRODUCTION --- p.2 / Chapter 1.1 --- rIbosome-inactivatIng proteins (RIPS) --- p.2 / Chapter 1.1.1 --- Classification of RIPs --- p.2 / Chapter 1.1.2 --- Distribution of RIPs --- p.6 / Chapter 1.1.3 --- Molecular biology of RIPs --- p.7 / Chapter 1.1.4 --- Physical and chemical properties of RIPs --- p.9 / Chapter 1.1.5 --- Enzymatic and translation-inhibitory activities --- p.12 / Chapter 1.1.6 --- RIP-based Immunotoxins --- p.16 / Chapter 1.2 --- MOMORDICA CHARANTIA and its RIBoosome-inactivating proteins (RIP) --- p.17 / Chapter 1.2.1 --- Momordica charantia --- p.17 / Chapter 1.2.2 --- Ribosome-inactivating proteins (RIPs) in Momordica charantia --- p.18 / Chapter 1.3 --- Objective of this study --- p.28 / Chapter CHAPTER 2 --- STUDY ON A NEW RIBOSOME-INACTIVATING PROTEIN (RIP) FROM MOMORDICA CHARANTIA SEEDS --- p.30 / Chapter 2.1 --- Introduction --- p.30 / Chapter 2.2 --- Materials and Methods --- p.33 / Chapter 2.2.1 --- Materials --- p.33 / Chapter 2.2.2 --- RIP isolation --- p.34 / Chapter 2.2.3 --- Characterization --- p.35 / Chapter 2.3 --- Results --- p.42 / Chapter 2.4 --- Discussion --- p.48 / Chapter CHAPTER 3 --- STUDY ON A NEW RIBOSOME-INACTIVATING PROTEIN (RIP) FROM MOMORDICA CHARANTIA FRUITS --- p.51 / Chapter 3.1 --- Introduction --- p.51 / Chapter 3.2 --- Materials and methods --- p.53 / Chapter 3.2.1 --- Materials --- p.53 / Chapter 3.2.2 --- RIP isolation --- p.54 / Chapter 3.2.3 --- Characterization --- p.56 / Chapter 3.3 --- Results --- p.56 / Chapter 3.4 --- Discussion --- p.62 / Chapter CHAPTER 4 --- GENERAL DISCUSSION AND CONCLUSION --- p.64 / Chapter 4.1 --- General discussion --- p.64 / Chapter 4.2 --- Conclusion --- p.72 / REFERENCES --- p.74

Typescript. Bibliography: leaves 51-61.

Graduation date: 2001

Thesis (Ph. D.)--University of Texas at Austin, 1999. / Vita. Includes bibliographical references (leaves 102-112). Available also in a digital version from Dissertation Abstracts.

published_or_final_version / Chemistry / Master / Master of Philosophy

No description available.

No description available.

This work was devoted to the study of (a) the physico-chemical, functional, and structural properties of bean (Phaseolus sp.) protein isolates in relation to their microstructures, and (b) the effects of protein carbohydrate interactions on physico-chemical, functional, and structural properties. The contents of protein, and both total and individual sugars of alkali extracted (amorphous) and citric acid extracted (bipyramidal and spheroidal) proteins from Phaseolus vulgaris (white kidney and navy) and Phaseolus lutanus (baby lima and large lima) beans were determined. The proteins were subjected to differential scanning calorimetry, and measurements of surface tension (air-water interface), surface hydrophobicity, and foam expansion. Structural analysis of the proteins were performed using Fourier transform infrared (FT-IR) spectroscopy. Enzymatic and chemical deglycosylation was performed on a white kidney bean protein isolate. / Glucose and mannose were the major sugars found in the isolates. Bipyramidal and spheroidal microstructures with higher protein contents generally had greater mannose content and lower glucose content. Differences in enthalpy of denaturation $( Delta$H), surface tension decay curves, surface hydrophobicities, and foam expansions were observed with isolates of different microstructures. Corresponding differences in molecular structure were not, however, detected by FT-IR spectroscopy. Using statistical analysis, a relationship between foam expansion and the $ Delta$H, solubility, surface hydrophobicity and surface tension of the isolates was obtained. Preliminary results suggest that the removal of carbohydrate influenced the physico-chemical properties of the protein.