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Serial analysis of gene expression during mycelium and primordium stages of shiitake mushroom (xianggu) Lentinula edodes.

Chum Wing Yan, Winnie. / Thesis submitted in 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 157-178). / Abstracts in English and Chinese. / English Abstract --- p.i / Chinese Abstract --- p.iii / Acknowledgements --- p.v / Abbreviations --- p.vi / Table of Contents --- p.vii / List of Figures --- p.x / List of Tables --- p.xiii / Chapter Chapter One --- Literature Review / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Life cycle --- p.2 / Chapter 1.3 --- Nutritional value --- p.4 / Chapter 1.4 --- Medicinal value --- p.4 / Chapter 1.4.1 --- Antitumor ability --- p.5 / Chapter 1.4.2 --- Antimicrobial ability --- p.5 / Chapter 1.4.3 --- Hypocholesterolaemic effect --- p.6 / Chapter 1.4.4 --- Anti-viral effect --- p.6 / Chapter 1.4.5 --- Anticaries effects --- p.7 / Chapter 1.6 --- Commercial value --- p.7 / Chapter 1.6.1 --- Biodecolorization --- p.7 / Chapter 1.6.2 --- Bioconversion --- p.7 / Chapter 1.6.3 --- Biodegradation --- p.8 / Chapter 1.6.4 --- Indicator --- p.9 / Chapter 1.7 --- Cultivation --- p.9 / Chapter 1.8 --- Content --- p.10 / Chapter 1.9 --- Transformation --- p.11 / Chapter 1.10 --- Gene regulation for growth and fruiting body development --- p.13 / Chapter 1.11 --- Serial Analysis of Gene Expression --- p.18 / Chapter 1.11.1 --- Introduction --- p.18 / Chapter 1.11.2 --- Principles --- p.20 / Chapter 1.11.3 --- SAGE Application in Cancer and Immunology Studies --- p.22 / Chapter 1.11.4 --- Improvement of SAGE --- p.23 / Chapter 1.11.5 --- Bioinformatics --- p.24 / Chapter 1.12 --- DNA Microarray --- p.27 / Chapter 1.12.1 --- Introduction --- p.27 / Chapter 1.12.2 --- Application --- p.28 / Chapter 1.12.3 --- Method of cDNA Microarray --- p.28 / Chapter Chapter Two --- Serial Analysis of Gene Expression / Chapter 2.1 --- Introduction --- p.30 / Chapter 2.2 --- Material and Methods --- p.32 / Chapter 2.2.1 --- Mushroom cultivation and RNA extraction --- p.32 / Chapter 2.2.2 --- RNA Quality Estimation --- p.33 / Chapter 2.2.3 --- mRNA Isolation --- p.34 / Chapter 2.2.4 --- Serial Analysis of Gene Analysis (SAGE) --- p.34 / Chapter 2.2.4.1 --- Binding mRNA to magnetic beads for cDNA synthesis --- p.34 / Chapter 2.2.4.2 --- DNA synthesis verification --- p.35 / Chapter 2.2.4.3 --- NlaIII digestion --- p.36 / Chapter 2.2.4.4 --- NlaIII digestion verification --- p.36 / Chapter 2.2.4.5 --- Adapters ligation --- p.37 / Chapter 2.2.4.6 --- Cleaving with tagging enzyme --- p.37 / Chapter 2.2.4.7 --- Ditags creation --- p.38 / Chapter 2.2.4.8 --- PCR optimization and scale-up --- p.38 / Chapter 2.2.4.9 --- Polyacrylamide gel electrophoresis --- p.39 / Chapter 2.2.4.10 --- Eluting DNA from the gel --- p.40 / Chapter 2.2.4.11 --- NlaIII Cleavage and polyacrylamide gel electrophoresis --- p.40 / Chapter 2.2.4.12 --- Concatemers Ligation --- p.41 / Chapter 2.2.4.13 --- Cloning Concatemers into pZErO®-1 --- p.42 / Chapter 2.2.5 --- One Shot® TOP 10 Electrocomp´ёØ E. Coli transformation --- p.42 / Chapter 2.2.6 --- PCR Screening and Sequencing --- p.44 / Chapter 2.2.7 --- Sequence Analysis --- p.44 / Chapter 2.3 --- Results --- p.45 / Chapter 2.3.1 --- RNA Extraction --- p.45 / Chapter 2.3.2 --- cDNA Synthesis --- p.45 / Chapter 2.3.3 --- NlaIII digestion --- p.45 / Chapter 2.3.4 --- PCR amplification --- p.46 / Chapter 2.3.5 --- Gel-purification of the 100bp Ditags --- p.46 / Chapter 2.3.6 --- Isolation of the 26bp Ditags --- p.46 / Chapter 2.3.7 --- Concatemers Generation --- p.47 / Chapter 2.3.8 --- PCR Screening --- p.47 / Chapter 2.3.9 --- The Abundance and Identity of SAGE Tags --- p.48 / Chapter 2.4 --- Discussion --- p.86 / Chapter 2.4.1 --- RNA Extraction --- p.86 / Chapter 2.4.2 --- cDNA Synthesis and NlaIII digestion --- p.86 / Chapter 2.4.3 --- PCP amplification --- p.87 / Chapter 2.4.4 --- SAGE Tags Analysis --- p.88 / Chapter Chapter Three --- Microarray / Chapter 3.1 --- Introduction --- p.96 / Chapter 3.2 --- Materials and Methods --- p.97 / Chapter 3.2.1 --- Microarray chip preparation --- p.97 / Chapter 3.3.2 --- Sample preparation --- p.97 / Chapter 3.2.3 --- cDNA Synthesis and Sample labeling --- p.98 / Chapter 3.2.4 --- cDNA Purification --- p.99 / Chapter 3.2.5 --- cDNA analysis --- p.99 / Chapter 3.2.6 --- Array Hybridization --- p.102 / Chapter 3.2.6.1 --- Sample Preparation --- p.102 / Chapter 3.2.6.2 --- Hybridization Procedure --- p.102 / Chapter 3.2.7 --- Stringency Washes --- p.103 / Chapter 3.2.8 --- Detection with TSA --- p.103 / Chapter 3.2.9 --- Scanning and Analysis --- p.105 / Chapter 3.3 --- Results --- p.109 / Chapter 3.4 --- Discussion --- p.120 / Chapter Chapter Four --- Full Length Sequencing / Chapter 4.1 --- Introduction --- p.124 / Chapter 4.2 --- Material and Methods --- p.124 / Chapter 4.3 --- Results and Discussion --- p.125 / Chapter Chapter Five --- General Discussion --- p.149 / Appendix --- p.155 / References --- p.157

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324447
Date January 2003
ContributorsChum, Wing Yan., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xiii, 178 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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