Cheng, Chi Keung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 144-160). / Abstracts in English and Chinese. / English Abstract --- p.ii / Chinese Abstract --- p.iv / Acknowledgements --- p.v / Abbreviations --- p.vi / Table of Contents --- p.vii / List of Figures --- p.x / List of Tables --- p.xii / Chapter Chapter 1 --- Literature Review / Chapter 1.1 --- Introduction & Taxonomy --- p.1 / Chapter 1.2 --- Life cycle and morphology --- p.1 / Chapter 1.3 --- Growth requirements --- p.4 / Chapter 1.3.1 --- Nutritional requirements --- p.4 / Chapter 1.3.2 --- Environment factors --- p.5 / Chapter 1.4 --- Fruiting body development in Coprinopsis cinerea --- p.6 / Chapter 1.4.1 --- Physiology of the fruiting process --- p.6 / Chapter 1.4.2 --- Other studies related to the fruiting process --- p.7 / Chapter 1.5 --- Other biological studies in Coprinopsis cinerea --- p.8 / Chapter 1.5.1 --- Meiosis studies --- p.9 / Chapter 1.5.2 --- Mating analyses --- p.10 / Chapter 1.5.3 --- Peroxidase production --- p.11 / Chapter 1.5.4 --- Transformation and gene silencing --- p.12 / Chapter 1.5.5 --- Other studies --- p.13 / Chapter 1.6 --- C. cinerea genome project --- p.13 / Chapter 1.7 --- Transcriptome analyses --- p.14 / Chapter 1.7.1 --- Serial Analysis of Gene Expression (SAGE) --- p.14 / Chapter 1.7.2 --- Analyzing the 5´ةend of transcripts --- p.16 / Chapter 1.7.3 --- Mapping of SAGE tags to the genome --- p.19 / Chapter 1.8 --- High throughput sequencing --- p.20 / Chapter 1.8.1 --- Pyrophosphate sequencing --- p.20 / Chapter 1.8.2 --- Application of pyrosequencing --- p.21 / Chapter 1.9 --- Aims of project --- p.22 / Chapter Chapter 2 --- 5' Serial Analysis of Gene Expression (5' SAGE) from mycelial and primordial stages of C. cinerea / Chapter 2.1 --- Introduction --- p.24 / Chapter 2.2 --- Materials and Methods --- p.29 / Chapter 2.2.1 --- 5' SAGE libraries construction --- p.29 / Chapter 2.2.1.1 --- Mushroom mycelium and primordium cultivation --- p.29 / Chapter 2.2.1.2 --- RNA extraction --- p.29 / Chapter 2.2.1.3 --- Isolation of mRNA --- p.30 / Chapter 2.2.1.4 --- cDNA synthesis --- p.31 / Chapter 2.2.1.5 --- Mmel digestion and Polyacrylamide gel electrophoresis --- p.32 / Chapter 2.2.1.6 --- Formation and amplification of ditag --- p.33 / Chapter 2.2.2 --- Identification of lOObp ditag --- p.34 / Chapter 2.2.3 --- High throughput pyrosequencing --- p.35 / Chapter 2.2.4 --- Tags extraction from ditags --- p.35 / Chapter 2.2.5 --- Genome mapping and annotation --- p.36 / Chapter 2.3 --- Results --- p.37 / Chapter 2.3.1 --- 5´ةSAGE libraries construction --- p.37 / Chapter 2.3.1.1 --- cDNA synthesis --- p.37 / Chapter 2.3.1.2 --- Mmel digestion and ditag formation --- p.38 / Chapter 2.3.2 --- Identification of lOObp ditags --- p.39 / Chapter 2.3.3 --- High throughput pyrosequencing --- p.40 / Chapter 2.3.4 --- Tags extraction from ditags --- p.41 / Chapter 2.3.5 --- Genome mapping and annotation --- p.42 / Chapter 2.4 --- Discussion --- p.46 / Chapter 2.4.1 --- 5´ةSAGE libraries construction --- p.46 / Chapter 2.4.2 --- Tags extraction and genome mapping --- p.46 / Chapter 2.4.3 --- Observations based on the genome mapping data --- p.48 / Chapter Chapter 3 --- Validation of expression patterns of 5' SAGE libraries and analysis of differentially expressed genes / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Materials and Methods --- p.58 / Chapter 3.2.1 --- Identification of housekeeping gene by Northern Blot analysis --- p.58 / Chapter 3.2.1.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.58 / Chapter 3.2.1.2 --- Transfer of RNAs --- p.58 / Chapter 3.2.1.3 --- Probe preparation --- p.59 / Chapter 3.2.1.4 --- "Hybridization, Stringency washes and signal detection" --- p.60 / Chapter 3.2.2 --- Quantitative real-time PCR --- p.61 / Chapter 3.2.2.1 --- cDNA synthesis from 2 developmental stages --- p.61 / Chapter 3.2.2.2 --- Primer design and verification --- p.62 / Chapter 3.2.2.3 --- Real time PCR reaction and data analysis --- p.65 / Chapter 3.2.3 --- Gene expression level comparison --- p.65 / Chapter 3.3 --- Results --- p.67 / Chapter 3.3.1 --- Identification of housekeeping gene by Northern Blot analysis --- p.67 / Chapter 3.3.2 --- Quantitative real-time PCR analysis --- p.71 / Chapter 3.3.3 --- Gene expression level comparison --- p.78 / Chapter 3.4 --- Discussion --- p.126 / Chapter 3.4.1 --- Validation of 5´ة SAGE libraries --- p.126 / Chapter 3.4.2 --- Analysis of highly and differentially expressed genes --- p.127 / Chapter Chapter 4 --- General discussion --- p.135 / References --- p.144 / Appendix --- p.161
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326258 |
| Date | January 2008 |
| Contributors | Cheng, Chi Keung., Chinese University of Hong Kong Graduate School. Division of Molecular Biotechnology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xii, 166 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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