Lo Ting Sze. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 145-170). / Abstracts in English and Chinese. / Declaration --- p.i / Abstract --- p.ii / Acknowledgements --- p.vii / List of Contents / Contents in Brief --- p.viii / Contents in Detail --- p.ix / List of Figures and Tables / List of Figures --- p.xiii / List of Tables --- p.xvi / Chapter Chapter 1 --- Introduction and literature review General introduction --- p.1 / Chapter 1.1 --- Structural changes in ovary of penaeid shrimp during maturation --- p.3 / Chapter 1.2 --- Biochemical and physiological changes in ovary of penaeid shrimp during maturation --- p.7 / Chapter 1.2.1 --- Variation of protein content --- p.7 / Chapter 1.2.1.1 --- Changes of proteins in ovary of penaeid shrimp during maturation --- p.7 / Chapter 1.2.1.2 --- Biochemical characterization of vitellin --- p.8 / Chapter 1.2.1.3 --- Site of vitellogenin synthesis --- p.10 / Chapter 1.2.2 --- Variation of lipid composition --- p.12 / Chapter 1.2.2.1 --- Role of lipids in ovary of penaeid shrimp --- p.12 / Chapter 1.2.2.2 --- Changes of lipids in ovary of penaeid shrimp during maturation --- p.13 / Chapter 1.2.3 --- Variation of other nutrient contents --- p.16 / Chapter 1.3 --- Endocrine control of ovarian maturation in penaeid shrimp --- p.16 / Chapter 1.3.1 --- Gonad-inhibiting hormone (GIH) --- p.17 / Chapter 1.3.2 --- Crustacean hyperglycemic hormone (CHH) --- p.19 / Chapter 1.3.3 --- X-organ sinus gland complex (XOSG) and CHH neuropeptide family --- p.20 / Chapter 1.3.4 --- Gonad-stimulating hormone (GSH) --- p.22 / Chapter 1.3.5 --- Methyl farnesoate (MF) --- p.23 / Chapter 1.3.6 --- Neurotransmitters --- p.24 / Chapter 1.3.7 --- Other factors --- p.26 / Chapter 1.3.8 --- Androgenic hormone (AH) --- p.27 / Chapter 1.4 --- Ecology and reproductive biology of Metapenaeus ensis --- p.29 / Chapter 1.4.1 --- Identification --- p.29 / Chapter 1.4.2 --- Ecology --- p.29 / Chapter 1.4.3 --- Reproductive biology --- p.30 / Chapter 1.5 --- Objective of research --- p.32 / Chapter Chapter 2 --- "Variation of circulating gonad-inhibiting hormone (GIH) level during ovarian maturation in the shrimp, Metapenaeus ensis" / Chapter 2.1 --- Introduction --- p.34 / Chapter 2.2 --- Materials and methods --- p.35 / Chapter 2.2.1 --- Experimental animals and serum extraction --- p.35 / Chapter 2.2.2 --- Protein content assay --- p.36 / Chapter 2.2.3 --- SDS-polyacrymide gel electrophoresis (SDS-PAGE) --- p.39 / Chapter 2.2.4 --- Western blot --- p.40 / Chapter 2.2.5 --- Purification of IgG from antiserum --- p.42 / Chapter 2.2.6 --- ELISA assay (indirect ELISA) --- p.42 / Chapter 2.3 --- Results --- p.44 / Chapter 2.3.1 --- Experimental animals --- p.44 / Chapter 2.3.2 --- Purification of anti-GIH IgG --- p.46 / Chapter 2.3.3 --- Optimization of indirect ELISA --- p.46 / Chapter 2.3.4 --- Determination of circulating GIH in haemolymph --- p.51 / Chapter 2.4 --- Discussion --- p.55 / Chapter Chapter 3 --- "Identification of genes differentially expressed during ovarian maturation in the shrimp, Metapenaeus ensis" / Chapter 3.1 --- Introduction --- p.60 / Chapter 3.2 --- Materials and methods --- p.61 / Chapter 3.2.1 --- Outline of methodology --- p.61 / Chapter 3.2.2 --- Experimental animals --- p.62 / Chapter 3.2.3 --- Total RNA extraction --- p.62 / Chapter 3.2.4 --- RNA arbitrarily primed PCR (RAP-PCR) --- p.64 / Chapter 3.2.5 --- DNA electrophoresis by agarose gel --- p.68 / Chapter 3.2.6 --- DNA electrophoresis by polyacrylamide gel --- p.68 / Chapter 3.2.7 --- Preparation of DIG-labeled probe from PCR product --- p.69 / Chapter 3.2.8 --- Checking for DIG-labeling yield --- p.69 / Chapter 3.2.9 --- Excision of cDNA library from ovary of Metapenaeus ensis --- p.70 / Chapter 3.2.10 --- PCR screening of insertion sequence --- p.73 / Chapter 3.2.11 --- Dot-blot --- p.75 / Chapter 3.2.12 --- Dot-blot hybridization --- p.76 / Chapter 3.2.13 --- Growth of cell for plasmid preparation --- p.79 / Chapter 3.2.14 --- Permanent culture preparation --- p.80 / Chapter 3.2.15 --- DNA Sequencing --- p.80 / Chapter 3.2.16 --- RNA formaldehyde denaturing gel electrophoresis --- p.82 / Chapter 3.2.17 --- Northern blot --- p.84 / Chapter 3.2.18 --- Northern hybridization --- p.84 / Chapter 3.2.19 --- 5´ة RACE (rapid amplification of cDNA ends) --- p.88 / Chapter 3.3 --- Results --- p.92 / Chapter 3.3.1 --- Experimental animals --- p.92 / Chapter 3.3.2 --- Total RNA extraction --- p.92 / Chapter 3.3.3 --- RNA arbitrarily primed PCR (RAP-PCR) --- p.92 / Chapter 3.3.4 --- Titer of cDNA library from Metapenaeus ensis ovary --- p.95 / Chapter 3.3.5 --- PCR screening of insertion sequence --- p.95 / Chapter 3.3.6 --- Dot-blot hybridization --- p.99 / Chapter 3.3.7 --- DNA sequencing of differentially expressed genes --- p.96 / Chapter 3.3.8 --- Northern analysis --- p.99 / Chapter 3.3.8.1 --- Housekeeping gene - elongation factor 1α --- p.106 / Chapter 3.3.8.2 --- Translationally controlled tumor protein (TCTP) --- p.106 / Chapter 3.3.8.3 --- Cytoskeletal actin --- p.109 / Chapter 3.3.8.4 --- Keratin --- p.109 / Chapter 3.3.8.5 --- Heat shock cognate 70 kD protein (hsc70) --- p.109 / Chapter 3.3.8.6 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.117 / Chapter 3.3.8.7 --- Arginine kinase --- p.117 / Chapter 3.3.8.8 --- High mobility group 1-like protein (HMGl-like protein) --- p.122 / Chapter 3.3.8.9 --- Nucleoside diphosphate kinase --- p.122 / Chapter 3.4 --- Discussion --- p.122 / Chapter 3.4.1 --- RAP-PCR and dot-blot hybridization --- p.122 / Chapter 3.4.2 --- Functions of differentially expressed genes in ovarian maturation --- p.128 / Chapter 3.4.2.1 --- Translationally controlled tumor protein (TCTP) --- p.128 / Chapter 3.4.2.2 --- Cytoskeletal actin --- p.130 / Chapter 3.4.2.3 --- Keratin --- p.130 / Chapter 3.4.2.4 --- Heat shock cognate 70 kD protein (hsc70) --- p.131 / Chapter 3.4.2.5 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.134 / Chapter 3.4.2.6 --- Arginine kinase --- p.135 / Chapter 3.4.2.7 --- High mobility group 1-like protein (HMGl-like protein) --- p.136 / Chapter 3.4.2.8 --- Nucleoside diphosphate kinase --- p.137 / Chapter 3.4.3 --- Speculation on the molecular changes in ooctyes during ovarian maturation --- p.139 / Chapter Chapter 4 --- General conclusion --- p.142 / References --- p.145
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324395 |
| Date | January 2003 |
| Contributors | Lo, Ting Sze., 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, xvi, 170 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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