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EST expression and proteomic studies on mycelia of cordyceps militaris.

Chan Ching-Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 109-123). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgements --- p.vi / Abbreviations --- p.vii / Table of contents --- p.xi / List of figures --- p.xiv / List of tables --- p.xv / Chapter 1. --- Introduction --- p.1 / Chapter 2. --- Literature review --- p.3 / Chapter 2.1 --- History --- p.3 / Chapter 2.2 --- The living environment and life cycles of Cordyccps --- p.3 / Chapter 2.3 --- Chemical constituents of Cordyceps --- p.4 / Chapter 2.3.1 --- Determintaion of active ingredients in Cordyceps --- p.6 / Chapter 2.4 --- Therapeutic Functions --- p.8 / Chapter 2.4.1 --- Cardiovascular and circulatory functions --- p.8 / Chapter 2.4.1.1 --- Effects on Cholesterol and lipid metabolism --- p.8 / Chapter 2.4.1.2 --- Dilation of vasculature and cerebolature --- p.8 / Chapter 2.4.2 --- Respiratory functions --- p.9 / Chapter 2.4.3 --- Renai functions --- p.9 / Chapter 2.4.3.1 --- Effccts on chronic renal failure patients --- p.9 / Chapter 2.4.3.2 --- Protective effects on kidney toxicity --- p.9 / Chapter 2.4.4 --- Hepatic functions --- p.10 / Chapter 2.4.4.1 --- Effect on hepatitis B patients --- p.10 / Chapter 2.4.4.2 --- Energy state of liver --- p.10 / Chapter 2.4.5 --- Aging and senescence: Longevity enhancement --- p.11 / Chapter 2.4.5.1 --- Senescence --- p.11 / Chapter 2.4.5.2 --- Antioxidant effects --- p.11 / Chapter 2.4.6 --- Immune functions --- p.12 / Chapter 2.4.6.1 --- Enhancing immune system --- p.12 / Chapter 2.4.6.2 --- Anti-tumor effects --- p.12 / Chapter 2.4.7 --- Reproductive effects --- p.13 / Chapter 2.4.8 --- Hyperglycemic effects --- p.13 / Chapter 2.5 --- Cultivation --- p.15 / Chapter 2.5.1 --- Carbon and nitrogen sources --- p.15 / Chapter 2.5.2 --- Initial pH and temperature --- p.16 / Chapter 2.5.3 --- Bioelements --- p.16 / Chapter 2.5.4 --- Agitation intensity --- p.16 / Chapter 2.5.5 --- Aeration rate --- p.18 / Chapter 2.6 --- Fungal genetics --- p.19 / Chapter 2.6.1 --- EST approach --- p.19 / Chapter 2.7 --- Proteomic studies --- p.21 / Chapter 2.7.1 --- 2D gel electrophoresis --- p.21 / Chapter 2.7.2 --- Mass spectrometry --- p.22 / Chapter 2.7.3 --- Limitations and improvements --- p.22 / Chapter 2.7.4 --- Multiple spots for the same proteins --- p.24 / Chapter 2.7.5 --- Fungal proteomics --- p.24 / Chapter 2.7.5.1 --- Extraction method --- p.24 / Chapter 2.7.5.2 --- Combined uses of EST sequences and amino acid sequences --- p.26 / Chapter 2.7.5.3 --- Glycosylation --- p.26 / Chapter 3. --- Materials and methods --- p.28 / Chapter 3.1 --- Genomic Studies --- p.28 / Chapter 3.1.1 --- Strains and growth conditions --- p.28 / Chapter 3.1.2 --- Total RNA Extraction --- p.28 / Chapter 3.1.3 --- Isolation of mRNA --- p.30 / Chapter 3.1.4 --- cDNA Library Construction --- p.30 / Chapter 3.1.5 --- PCR Screening --- p.31 / Chapter 3.1.6 --- EST sequencing --- p.31 / Chapter 3.1.7 --- EST assembling and annotation --- p.32 / Chapter 3.2 --- Proteomic Studies --- p.33 / Chapter 3.2.1 --- Sample Preparation --- p.33 / Chapter 3.2.2 --- Quantitation --- p.34 / Chapter 3.2.3 --- 2-D PAGE --- p.34 / Chapter 3.2.4 --- In-gel digestion and peptide extraction --- p.36 / Chapter 3.2.5 --- MALDI-TOF MS Analysis --- p.37 / Chapter 3.3 --- Determination of adenosine using RP-HPLC --- p.38 / Chapter 4. --- Result --- p.39 / Chapter 4.1 --- Genomic studies --- p.39 / Chapter 4.1.1 --- cDNA library --- p.39 / Chapter 4.1.2 --- cDNA sequence analysis --- p.39 / Chapter 4.1.3 --- Functional annotation and analysis --- p.42 / Chapter 4.2 --- Proteomic --- p.67 / Chapter 4.2.1 --- 2D analysis and resolution --- p.67 / Chapter 4.2.2 --- Protein identification and annotation --- p.74 / Chapter 4.2.3 --- Image analysis --- p.81 / Chapter 4.3 --- Presence of adenosine (HPLC) --- p.84 / Chapter 5. --- Discussion and conclusion --- p.91 / Chapter 5.1 --- Genomic studies --- p.91 / Chapter 5.2 --- Presence of adenosine --- p.95 / Chapter 5.3 --- Proteomic --- p.96 / Chapter 5.3.1 --- Protein with increasing expression level --- p.97 / Chapter 5.3.1.1 --- MEI5 (Spot1084) --- p.97 / Chapter 5.3.1.2 --- Hsp70 and hsp60 (Spot 894 & 903) --- p.97 / Chapter 5.3.1.3 --- GRP 78 (Spot 1085) --- p.98 / Chapter 5.3.1.4 --- Ubiquitin (Spot1071) --- p.98 / Chapter 5.3.1.5 --- "Serine-tRNA ligase, glutaminyl-tRNA synthase (Spot 1037, 924)" --- p.99 / Chapter 5.3.1.6 --- 2-isopropylmalate synthase (Spot 862) --- p.99 / Chapter 5.3.1.7 --- Acyl-CoA oxidase 3 (Spot 882) --- p.100 / Chapter 5.3.1.8 --- ATP synthase beta chain (Spot 937) --- p.100 / Chapter 5.3.2 --- Proteins with decreasing expression --- p.101 / Chapter 5.3.2.1 --- 14-3-3 protein (spot 1080) --- p.101 / Chapter 5.3.2.2 --- Actin (Spot 945) --- p.101 / Chapter 5.3.2.3 --- GTP binding protein SPI1 (Spot1031) --- p.102 / Chapter 5.3.2.4 --- Hoclp (Spot 972) --- p.102 / Chapter 5.3.2.5 --- Rchl8p (Spot 983) --- p.103 / Chapter 5.3.2.6 --- Formaldehyde dehydrogenase (Spot 958) --- p.103 / Chapter 5.3.2.7 --- V-type ATPase (Spot 961) --- p.104 / Chapter 5.3.2.8 --- Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Spot 987) --- p.104 / Chapter 5.3.2.9 --- Idp3p (Spot 929) --- p.105 / References

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324986
Date January 2004
ContributorsChan, Ching-Man., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvi, 123 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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