Hui Cheung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 166-172). / Abstracts in English and Chinese. / Abstract --- p.iii / Acknowledgments --- p.vi / Abbreviations --- p.vii / Table of Contents --- p.viii / Table of Figures --- p.xiii / Table of Tables --- p.xviii / Chapter Chapter 1. --- Literature Reviews / Chapter 1.1 --- Functional Genomics and Its Developments --- p.1 / Chapter 1.1.1 --- Introduction --- p.1 / Chapter 1.1.2 --- "Transcriptomics, Proteomics and Metabolomics" --- p.1 / Chapter 1.1.3 --- Gene-perturbing Strategies --- p.3 / Chapter 1.1.4 --- Applications of Functional Genomics --- p.4 / Chapter 1.2 --- Serial Analysis of Gene Expression (SAGE) and Generation of Longer cDNA Fragments from SAGE tags for Gene Identification (GLGI) --- p.6 / Chapter 1.2.1 --- Introduction --- p.6 / Chapter 1.2.2 --- Principles and Methods of SAGE --- p.6 / Chapter 1.2.3 --- Data Analysis --- Bioinformatics --- p.9 / Chapter 1.2.4 --- Applications of SAGE --- p.9 / Chapter 1.2.5 --- Modifications of SAGE --- p.10 / Chapter 1.2.6 --- Principles and Methods of GLGI --- p.11 / Chapter 1.2.7 --- Applications and Improvements of GLGI --- p.14 / Chapter 1.3 --- Transformation --- p.15 / Chapter 1.3.1 --- Introduction --- p.15 / Chapter 1.3.2 --- Different Methods of Transformation --- p.15 / Chapter 1.3.2.1 --- General Transformation Strategy --- p.15 / Chapter 1.3.2.2 --- Polyethylene Glycol (PEG)-mediated Transformation --- p.16 / Chapter 1.3.2.3 --- Restriction Enzyme Mediated Integration (REMI) --- p.16 / Chapter 1.3.2.4 --- Electroporation --- p.17 / Chapter 1.3.2.5 --- Particle Bombardment --- p.17 / Chapter 1.3.3 --- The Future Needs of Transformation --- p.18 / Chapter 1.4 --- RNA Silencing --- p.20 / Chapter 1.4.1 --- Introduction --- p.20 / Chapter 1.4.2 --- Major Components and Principles of RNAi --- p.21 / Chapter 1.4.3 --- Applications of RNA Silencing --- p.23 / Chapter 1.5 --- The Target Organism Lentinula edodes --- p.25 / Chapter 1.5.1 --- Introduction --- p.25 / Chapter 1.5.2 --- The Life Cycle of L. edodes --- p.26 / Chapter 1.5.3 --- Biochemical and Molecular Studies on L. edodes --- p.27 / Chapter 1.5.4 --- Prospectus --- p.29 / Chapter Chapter 2. --- Development of Methods for Studying Gene Function in Lentinula edodes / Chapter 2.1 --- Introduction --- p.30 / Chapter 2.2 --- Materials and Methods --- p.32 / Chapter 2.2.1 --- Cultivation of Lentinula edodes --- p.32 / Chapter 2.2.2 --- Proplast Release and Regeneration --- p.32 / Chapter 2.2.3 --- Preparation of Plasmid DNA --- p.33 / Chapter 2.2.4 --- Selectable Marker …Bialaphos --- p.35 / Chapter 2.2.5 --- Transformation --- p.35 / Chapter 2.2.5.1 --- Electroporation --- p.35 / Chapter 2.2.5.2 --- PEG-mediated Transformation --- p.36 / Chapter 2.3 --- Results --- p.37 / Chapter 2.3.1 --- Cultivation of Lentinula edodes --- p.37 / Chapter 2.3.2 --- Proplast Release and Regeneration --- p.37 / Chapter 2.3.3 --- Preparation of Plasmid DNA --- p.43 / Chapter 2.3.4 --- Selectable Marker--- Bialaphos --- p.43 / Chapter 2.3.5 --- Transformation --- p.46 / Chapter 2.3.5.1 --- Electroporation --- p.46 / Chapter 2.3.5.2 --- PEG-mediated Transformation --- p.46 / Chapter 2.4 --- Discussions and Conclusions --- p.57 / Chapter Chapter 3. --- Identification of Interested Genes in Expression Profile of SAGE using GLGI Method. / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.1.1 --- Results of SAGE Analysis --- p.61 / Chapter 3.1.2 --- Use of GLGI Method for Extension of SAGE Tags --- p.63 / Chapter 3.1.3 --- 5´ة Extension of GLGI (5'GLGI) --- p.65 / Chapter 3.1.3.1 --- Introduction --- p.65 / Chapter 3.1.3.2 --- "Overall strategy of 5, GLGI Method" --- p.67 / Chapter 3.1.3.3 --- Two-Steps PCR Method --- p.69 / Chapter 3.2 --- Generation of Longer cDNA Fragments from SAGE tags for Gene Identification (GLGI) --- p.71 / Chapter 3.2.1 --- Materials and Methods (GLGI Analysis) --- p.71 / Chapter 3.2.1.1 --- Total RNA Extraction --- p.71 / Chapter 3.2.1.2 --- Messenger RNA (mRNA) Extraction --- p.72 / Chapter 3.2.1.3 --- Preparation of 3´ة cDNA for GLGI --- p.73 / Chapter 3.2.1.4 --- NIaIII digestion of double strand cDNA --- p.74 / Chapter 3.2.1.5 --- PCR amplification of the 3'-cDNAs (Optional) --- p.77 / Chapter 3.2.1.6 --- GLGI Amplification of The Target Template --- p.80 / Chapter 3.2.1.7 --- DNA Cloning (Optional) --- p.82 / Chapter 3.2.1.8 --- Sequencing of GLGI PCR products --- p.85 / Chapter 3.2.2 --- 5' Materials and Methods (5' GLGI Analysis) --- p.86 / Chapter 3.2.2.1 --- Preparation of unique antisense primers --- p.86 / Chapter 3.2.2.2 --- 5' extension of GLGI products --- p.87 / Chapter 3.2.2.3 --- DNA Cloning (Optional) --- p.89 / Chapter 3.2.2.4 --- Sequencing of 5' GLGI PCR products --- p.89 / Chapter 3.2.3 --- Results (GLGI Analysis) --- p.90 / Chapter 3.2.3.1 --- Total RNA Extraction --- p.90 / Chapter 3.2.3.2 --- Messenger RNA Extraction --- p.90 / Chapter 3.2.3.3 --- Preparation of 3' cDNA for GLGI --- p.90 / Chapter 3.2.3.4 --- NIaIII digestion of double strand cDNA --- p.94 / Chapter 3.2.3.5 --- GLGI Amplification of The Target Template --- p.94 / Chapter 3.2.3.6 --- Sequencing of GLGI PCR products --- p.103 / Chapter 3.2.4 --- Results (5' GLGI Analysis) --- p.111 / Chapter 3.2.4.1 --- 5' extension of GLGI products --- p.111 / Chapter 3.2.4.2 --- Sequencing of 5´ة GLGI PCR products --- p.116 / Chapter 3.3 --- Discussions and Conclusions --- p.126 / Chapter 3.3.1 --- GLGI amplification of the target template --- p.126 / Chapter 3.3.2 --- 5' extension of GLGI products --- p.129 / Chapter 3.3.3 --- Two-Steps PCR Method --- p.130 / Chapter 3.3.4 --- Sequencing results of GLGI method and 5' GLGI method --- p.131 / Chapter Chapter 4. --- Identification of Unknown EST Using PCR Method With cDNA Library / Chapter 4.1 --- Introduction --- p.134 / Chapter 4.2 --- Materials and Methods --- p.134 / Chapter 4.2.1 --- Extension of 5' end of EST sequence by PCR method --- p.134 / Chapter 4.2.2 --- Purification of PCR products --- p.136 / Chapter 4.2.3 --- Sequencing of Extended EST products --- p.136 / Chapter 4.3 --- Results --- p.137 / Chapter 4.3.1 --- Extension of 5' end of EST sequence by PCR method --- p.137 / Chapter 4.3.2 --- Sequencing of Extended EST products --- p.137 / Chapter 4.4 --- Discussions and Conclusions --- p.147 / Chapter Chapter 5. --- General Discussions --- p.151 / Appendix I --- p.156 / Reference --- p.166
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325247 |
| Date | January 2005 |
| Contributors | Hui, Cheung., Chinese University of Hong Kong Graduate School. Division of Biology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xviii, 172 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use 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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