by Sham Lok To. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 143-171). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgement --- p.iv / Abbreviations --- p.v / Table of contents --- p.vi / List of figures --- p.ix / List of tables --- p.xi / Chapter Chapter One --- Literature Review --- p.1 / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- About L. edodes --- p.3 / Chapter 1.1.2 --- Nutritional and medicinal values of L. edodes --- p.4 / Chapter 1.1.3 --- Life cycle of L. edodes --- p.6 / Chapter 1.1.4 --- Environmental factors affecting fruiting body formation in L. edodes --- p.6 / Chapter 1.2 --- Molecular mechanisms of fruiting body development in L. edodes --- p.8 / Chapter 1.2.1 --- Expression profiling and identification of differentially expressed genes during fruiting --- p.8 / Chapter 1.2.2 --- Changing in membrane structure --- p.11 / Chapter 1.2.3 --- The signal transduction cascade --- p.12 / Chapter 1.3 --- Transformation in L. edodes and in other fungi --- p.14 / Chapter 1.3.1 --- Transformation of L. edodes --- p.14 / Chapter 1.3.2 --- Transformation in other fungi --- p.17 / Chapter 1.4 --- Bioinformatics tools for comparative promoter analysis --- p.22 / Chapter 1.5 --- Objectives and significance --- p.26 / Chapter Chapter Two --- Promoter analysis of differentially expressed genes (DEGs) in the fruiting body development in L. edodes --- p.27 / Chapter 2.1 --- Introduction --- p.27 / Chapter 2.2 --- Materials and methods --- p.29 / Chapter 2.2.1 --- Strains and cultivation conditions --- p.29 / Chapter 2.2.2 --- Genome walking of the 5' flanking region of the DEGs --- p.29 / Chapter 2.2.3 --- Annealing Control Primed (ACP) PCR --- p.31 / Chapter 2.2.4 --- Construction of genomic DNA library --- p.36 / Chapter 2.2.5 --- Nested PCR to amplify the target sequences --- p.37 / Chapter 2.2.6 --- Cloning and sequencing of the 5' flanking region --- p.38 / Chapter 2.2.7 --- Determination of transcription start site by the Neural Network algorithm --- p.39 / Chapter 2.2.8 --- Identification of putative transcription factor binding sites --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Construction of adaptor linked template for genome walking --- p.41 / Chapter 2.3.2 --- Sequence analysis and quality control --- p.41 / Chapter 2.3.3 --- Comparison of various methods in genome walking --- p.42 / Chapter 2.3.4 --- Promoter analysis --- p.42 / Chapter 2.4 --- Discussion --- p.58 / Chapter Chapter Three --- In-silico analysis of transcription factor binding sites and identification transcription factors expressed in L. edodes --- p.64 / Chapter 3.1 --- Introduction --- p.64 / Chapter 3.2 --- Material and methods --- p.67 / Chapter 3.2.1 --- Sequence manipulation and extraction of homologous ESTs from C. cinereus --- p.67 / Chapter 3.2.2 --- Extraction of 5' flanking region of the corresponding ESTs and promoter prediction --- p.67 / Chapter 3.2.3 --- Positional cloning of mating type factor A --- p.68 / Chapter 3.3 --- Results --- p.70 / Chapter 3.3.1 --- Sequence extraction and manipulation --- p.70 / Chapter 3.3.2 --- In-silico analysis of transcription factor binding sites in C. cinereus . --- p.70 / Chapter 3.3.3 --- Comparison of putative TFBS between L. edodes and C. cinereus --- p.71 / Chapter 3.3.4 --- Identification of transcription factors in L. edodes by positional cloning --- p.71 / Chapter 3.4 --- Discussion --- p.85 / Chapter Chapter Four --- Identification，expression profiling and promoter analysis of hydrophobin genes --- p.91 / Chapter 4.1 --- Introduction --- p.91 / Chapter 4.2 --- Material and methods --- p.92 / Chapter 4.2.1 --- Clustering and grouping of the hydrophobin ESTs --- p.92 / Chapter 4.2.2 --- Identification of the consensus sequences of the hydrophobin groups --- p.93 / Chapter 4.2.3 --- RNA Sources and Preparation --- p.93 / Chapter 4.2.4 --- Expression profiling of hydrophobin genes by RT-PCR --- p.95 / Chapter 4.2.5 --- Promoter cloning and analysis of hydrophobin genes --- p.95 / Chapter 4.3 --- Results --- p.97 / Chapter 4.3.1 --- Isolation and characterization of four newly found hydrophobin genes --- p.97 / Chapter 4.3.2 --- Expression levels of hydrophobins --- p.100 / Chapter 4.3.3 --- Promoter sequencing of the hydrophobins --- p.103 / Chapter 4.4 --- Discussion --- p.103 / Chapter Chapter Five --- Transformation of L. edodes --- p.110 / Chapter 5.1 --- Introduction --- p.110 / Chapter 5.2 --- Materials and methods --- p.112 / Chapter 5.2.1 --- Vectors and primers design --- p.112 / Chapter 5.2.2 --- Maxi-preparation of plasmids --- p.112 / Chapter 5.2.3 --- Cultural condition and optimization of protoplasts release --- p.114 / Chapter 5.2.4 --- PEG mediated transformation --- p.115 / Chapter 5.2.5 --- Electroporation mediated transformation --- p.116 / Chapter 5.2.6 --- PCR screening of regenerated transformant --- p.116 / Chapter 5.2.7 --- Particle bombardment --- p.117 / Chapter 5.3 --- Results --- p.121 / Chapter 5.4 --- Discussion --- p.128 / Chapter Chapter Six --- General discussions --- p.132 / References --- p.143
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325666 |
| Date | January 2006 |
| Contributors | Sham, Lok To., 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, xii, 171 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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