Tjia Wai Mui. / Thesis submitted in: July 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 123-140). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Acknowledgement --- p.iv / Abbreviations --- p.v / Table of Contents --- p.vi / List of Figures --- p.x / List of Tables --- p.xii / Chapter Chapter One --- Literature Review / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Growth of L. edodes --- p.3 / Chapter 1.2.1 --- Life cycle of L. edodes --- p.3 / Chapter 1.2.2 --- Growth parameters of L. edodes --- p.6 / Chapter 1.2.2.1 --- Temperature --- p.6 / Chapter 1.2.2.2 --- Relative humidity --- p.7 / Chapter 1.2.2.3 --- Moisture content in substrate --- p.7 / Chapter 1.2.2.4 --- Light --- p.8 / Chapter 1.2.2.5 --- pH --- p.8 / Chapter 1.3 --- Cultivation of L. edodes --- p.9 / Chapter 1.3.1 --- History and development of artificial cultivation --- p.9 / Chapter 1.3.2 --- Use of forced fruiting --- p.11 / Chapter 1.4 --- Molecular studies of stress on fungi --- p.12 / Chapter 1.4.1 --- Studies of temperature stress in mushroom --- p.12 / Chapter 1.4.2 --- Studies of molecular chaperones in fungi --- p.13 / Chapter 1.4.2.1 --- Role of molecular chaperones --- p.13 / Chapter 1.4.2.2 --- Heat shock protein 70 (Hsp70) and their cochaperones --- p.13 / Chapter 1.4.2.3 --- Other chaperones --- p.15 / Chapter 1.4.2.4 --- Molecular chaperones and development --- p.16 / Chapter 1.5 --- Prospectus --- p.19 / Chapter Chapter Two --- The Effect of Stress on the Growth of L. edodes / Chapter 2.1 --- Introduction --- p.23 / Chapter 2.2 --- Materials and Methods --- p.24 / Chapter 2.2.1 --- Strain and culture conditions --- p.24 / Chapter 2.2.2 --- Stress treatments --- p.24 / Chapter 2.2.3 --- Data collection --- p.25 / Chapter 2.2.4 --- Data analysis --- p.25 / Chapter 2.3 --- Results --- p.27 / Chapter 2.3.1 --- Reliability analysis --- p.27 / Chapter 2.3.2 --- Descriptive statistics --- p.28 / Chapter 2.3.3 --- Independent t-test (ANOVA) --- p.33 / Chapter 2.4 --- Discussion --- p.37 / Chapter Chapter Three --- Sequence Analysis of selected Stress Genes / Chapter 3.1 --- Introduction --- p.39 / Chapter 3.2 --- Materials and Methods --- p.40 / Chapter 3.2.1 --- Isolation of stress genes --- p.40 / Chapter 3.2.1.1 --- Construction of primordial cDNA library --- p.40 / Chapter 3.2.1.2 --- Screening of cDNA clones --- p.40 / Chapter 3.2.2 --- Sequence analyses of stress genes --- p.41 / Chapter 3.2.2.1 --- Amplification and purification of cDNA insert --- p.41 / Chapter 3.2.2.2 --- Full length DNA cycle sequencing --- p.42 / Chapter 3.2.2.3 --- Sequence analyses --- p.43 / Chapter 3.2.3 --- Screening of LeSSA (Inducible HSP70) --- p.45 / Chapter 3.2.3.1 --- PCR screening of LeSSA by degenerate primers and LeSSB specific primers --- p.45 / Chapter 3.2.3.2 --- Screening of LeSSA from cDNA library by hybridization --- p.49 / Chapter 3.3 --- Results --- p.51 / Chapter 3.3.1 --- Sequence analyses --- p.51 / Chapter 3.3.1.1 --- LeSSB --- p.51 / Chapter 3.3.1.2 --- LeMge1 --- p.57 / Chapter 3.3.1.3 --- LeSTI1 --- p.62 / Chapter 3.3.1.4 --- LeTCP1β --- p.69 / Chapter 3.3.1.5 --- LeTCP1γ --- p.74 / Chapter 3.3.2 --- Failure of isolating LeSSA (Inducible HSP70) --- p.80 / Chapter 3.4 --- Discussion --- p.82 / Chapter 3.4.1 --- Sequence analyses --- p.82 / Chapter 3.4.2 --- Screening of LeSSA --- p.84 / Chapter Chapter Four --- Characterization of stress genes upon different stresses / Chapter 4.1 --- Introduction --- p.86 / Chapter 4.2 --- Materials and Methods --- p.87 / Chapter 4.2.1 --- Strain and culture conditions --- p.87 / Chapter 4.2.2 --- Stress treatments --- p.87 / Chapter 4.2.3 --- Isolation of total RNAs --- p.87 / Chapter 4.2.4 --- Reverse transcriptase-polymerase chain reaction (RT-PCR) --- p.88 / Chapter 4.2.4.1 --- Reverse transcription --- p.88 / Chapter 4.2.4.2 --- PCR amplification by specific primers of stress genes --- p.89 / Chapter 4.2.5 --- Northern blot analyses --- p.91 / Chapter 4.2.5.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.91 / Chapter 4.2.5.2 --- Northern blotting --- p.91 / Chapter 4.2.5.3 --- Preparation of probes --- p.92 / Chapter 4.2.5.4 --- Hybridization and stringency washes --- p.93 / Chapter 4.2.6 --- Isolation of total protein --- p.94 / Chapter 4.2.7 --- Quantification of protein by Bradford method --- p.95 / Chapter 4.2.8 --- Western blot analyses --- p.95 / Chapter 4.2.8.1 --- Sodium dodecyl sulfate ´ؤ polyacrylamide gel electrophoresis (SDS-PAGE) --- p.95 / Chapter 4.2.8.2 --- Western blotting --- p.96 / Chapter 4.2.8.3 --- Immunodetection --- p.98 / Chapter 4.2.8.4 --- ECL detection --- p.98 / Chapter 4.3 --- Results --- p.99 / Chapter 4.3.1 --- Reverse transcriptase-polymerase chain reaction (RT-PCR) --- p.99 / Chapter 4.3.2 --- Northern blot hybridization --- p.106 / Chapter 4.3.2.1 --- Establishing an internal control --- p.106 / Chapter 4.3.2.2 --- Dig-labelling of stress genes --- p.106 / Chapter 4.3.2.3 --- Northern blot hybridizaton of stress genes --- p.106 / Chapter 4.3.3 --- Western blot hybridization --- p.111 / Chapter 4.4 --- Discussions --- p.113 / Chapter Chapter Five --- General Discussions --- p.118 / References --- p.123
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324437 |
| Date | January 2003 |
| Contributors | Tjia, Wai Mui., 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, 140 leaves : ill. ; 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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