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Biological activities and molecular cloning of a novel mannose-binding lectin isolated from the orchid (Dendrobium nobile).

by Luk Choi Wan. / Thesis submitted in: November 2005. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 125-132). / Abstracts in English and Chinese. / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.viii / Tabel of contents --- p.x / List of Figures --- p.xii / List of Tables --- p.xiv / List of Abbreviations --- p.xv / List of Abbreviations --- p.xv / Chapter Chapter One --- Literature review --- p.1 / Chapter 1.1 --- General introduction --- p.1 / Chapter 1.2 --- General aspects of plant lectins --- p.4 / Chapter 1.3 --- Monocot mannose-binding lectins --- p.5 / Chapter 1.3.1 --- General introduction --- p.5 / Chapter 1.3.2 --- Sugar specificity --- p.6 / Chapter 1.3.3 --- Isolation and purification --- p.7 / Chapter 1.3.4 --- Molecular cloning --- p.9 / Chapter 1.3.5 --- Molecular structure and modifications --- p.10 / Chapter 1.3.6 --- Molecular evolution --- p.12 / Chapter 1.3.7 --- Transformation --- p.12 / Chapter 1.3.8 --- Physiological roles --- p.14 / Chapter 1.3.9 --- Application --- p.19 / Chapter 1.4 --- Dendrobium nobile --- p.31 / Chapter 1.4.1 --- Background --- p.31 / Chapter 1.4.2 --- Chemical analysis --- p.32 / Chapter Chapter Two --- Biological activities of a mannose-binding lectin isolated from Dendrobium mobile --- p.33 / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and methods --- p.35 / Chapter 2.2.1 --- Mannose-binding lectin from D. nobile --- p.35 / Chapter 2.3 --- Biological activities of mannose-binding lectin from D. nobile --- p.36 / Chapter 2.3.1 --- Hemagglutinating activity --- p.36 / Chapter 2.3.2 --- In vitro anti-proliferative assay --- p.37 / Chapter 2.3.3 --- In vitro antiviral assay --- p.40 / Chapter 2.3.4 --- Statistical Analysis --- p.42 / Chapter 2.4 --- Results --- p.43 / Chapter 2.4.1 --- Physiochemical properties of D. nobile lectins --- p.43 / Chapter 2.4.2 --- Cytotoxicity to cancer cell lines --- p.53 / Chapter 2.4.3 --- Antiviral activity --- p.59 / Chapter 2.5 --- Discussion --- p.60 / Chapter Chapter Three --- Molecular cloning of lectin gene of Dendrobium nobile --- p.65 / Chapter 3.1 --- Introduction --- p.65 / Chapter 3.2 --- Methods --- p.67 / Chapter 3.2.1 --- RNA extraction --- p.67 / Chapter 3.2.2 --- RT-PCR synthesis of D. nobile lectin cDNA --- p.68 / Chapter 3.2.3 --- RACE of D. nobile agglutinin gene --- p.69 / Chapter 3.2.4 --- Generation of DNL full-length lectin gene sequence --- p.70 / Chapter 3.2.5 --- Cloning and sequencing of PCR product --- p.71 / Chapter 3.2.6 --- Data analyses --- p.72 / Chapter 3.2.7 --- Synthesis of single-stranded DIG-labeled DNA probe --- p.74 / Chapter 3.2.8 --- Northern blot analysis --- p.74 / Chapter 3.2.9 --- Genomic DNA extraction --- p.75 / Chapter 3.2.10 --- Southern blot analysis --- p.76 / Chapter 3.2.11 --- Expression of DNL in E. coli --- p.76 / Chapter 3.2.12 --- Western blot analysis --- p.78 / Chapter 3.3 --- Results --- p.80 / Chapter 3.3.1 --- Isolation and characterization of DNL gene --- p.80 / Chapter 3.3.2 --- Sequence analysis of DNL --- p.89 / Chapter 3.2.3 --- Secondary and tertiary structure --- p.96 / Chapter 3.2.4 --- Southern blot analysis --- p.99 / Chapter 3.2.5 --- Northern blot analysis --- p.99 / Chapter 3.2.6 --- Expression of fusion protein in E.coli --- p.102 / Chapter 3.4 --- Discussion --- p.104 / Chapter Chapter Four --- General Discussion --- p.110 / Chapter 4.1 --- General discussion --- p.110 / Chapter 4.2 --- Isolation and Characterization of monocot mannose-binding lectin of D. nobile --- p.111 / Chapter 4.3 --- Molecular cloning of monocot mannose-binding lectin of D. nobile --- p.115 / Chapter 4.4 --- Further investigations --- p.122 / Chapter Chapter Five --- Conclusion --- p.124 / References: --- p.125 / Appendix --- p.133

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325468
Date January 2006
ContributorsLuk, Choi Wan., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, leaves : ill. ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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