D.Sc. (Zoology) / The development of the chondrocranium, including the procartilaginous as well as some mesenchymatous developmental stages, was described in the chinese grass carp, ctenopharyngodon idella (Val.). Fourteen developmental stages, ranging from 5,0 mm to 11,0 mm, were microtomized, stained with Heidenhain's azan solution, mounted and examined. Seven developmental stages were described using drawings and reconstructions made from frontal, sagittal and transverse sections. The ontogenesis of the skeletal structures in the ethmoidal, orbitotemporal and otico-occipital regions of the neurocranium was investigated. Special attention was given to the development of the cranial muscles as well as to the ontogenesis of the skeletal structures of the viscerocranium. The lamina precerebralis-, noticed in other cyprinids as well as in the siluroids, was regarded as the morphological anterior wall of the brain capsule and the posterior part of the nasal septum as the morphological posterior wall of the nasal capsule, greatly reduced in Teleosts. The lamina orbitonasalis represents a supra-pharyngopremandibular. In the orbitotemporal region the cartilaginous trabecula and polar cartilage are initially separated by a procartilaginous bridge which forms the side-wall of the carotid incisure. A pila lateralis is absent in Ctenopharyngodon idella. The prootic, chondrifies intramembranously, a fact which supports Bjerring's(1977) view that the otic capsules are derived from the dermatomes. The composition of the palatoquadrate and of the hyomandibular in elasmobranchs and teleosts are discussed. The development of the internal and external derivatives of the dorsal constrictors of the branchial arches were investigated and their possible homoloques in the prootic visceral arches discussed. The levator and adductor muscles of the opercle probably represent a modified musculus adductor arcus hyoidei.

Thesis (Ph. D.)--University of Washington, 1990. / Vita. Includes bibliographical references (leaves [161]-177).

Thesis (M.Phil.)--City University of Hong Kong, 2005. / "Submitted to Department of Biology and Chemistry in partial fulfillment of the requirements for the degree of Master of Philosophy" Includes bibliographical references (leaves 124-141)

In November, 1983, 93 sexually immature and 33 mature grass carp (Ctenopharyngodon idella Val.) were surgically sterilized by clipping the gonaduct or removing a section of the ovaries of testes through a ventral incision between the pelvic girdle and the vent. No mortality occurred from this procedure. After six months (May, 1984), 95.7% of the immature and 100% of the mature grass carp had at least some gonadal tissue regeneration. Fifty-nine percent of the immature fish regenerated a pathway for the eggs or milt to enter the environment. Artificial spawning of the mature female grass carp was 66.7% successful. Over 20,000 fry resulted from the four females that ovulated. Due to the rapid regeneration of functional reproductive tissues, and the extrusion of the viable eggs by surgically altered fish, surgical sterilization was determined to be impractical for producing sterile grass carp for weed control.

by Wong Mee-wa. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 100-104. / Contents / Abstract / Abbreviations / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Biological Functions and Structure of GH --- p.1 / Chapter 1.3 --- Molecular Cloning of GH cDNA and its Expression --- p.3 / Chapter 1.4 --- Molecular Cloning of the Genomic Sequence of Growth Hormone --- p.5 / Chapter 1.4.1 --- The Isolation of Mammalian GH gene --- p.5 / Chapter 1.4.2 --- Molecular Cloning of Rainbow Trout Growth Hormone Gene --- p.6 / Chapter 1.4.3 --- Molecular Cloning of Atlantic Salmon Growth Hormone Gene --- p.9 / Chapter 1.4.4 --- Molecular Evolution of GH gene --- p.9 / Chapter 1.4.5 --- Control Elements on GH gene --- p.12 / Chapter 1.4.6 --- Production of Transgenic Fish --- p.14 / Chapter 1.5 --- Purpose of present Study --- p.15 / Chapter Chapter 2 --- Construction of a Genomic Library --- p.17 / Chapter 2.1 --- Introduction --- p.17 / Chapter 2.2 --- Strategy --- p.19 / Chapter 2.3 --- Materials and Methods --- p.20 / Chapter 2.3.1 --- Materials --- p.20 / Chapter 2.3.2 --- Procedure --- p.24 / Chapter 2.3.2.1 --- Extraction of Total Genomic DMA --- p.24 / Chapter 2.3.2.2 --- Southern Blotting and Hybridization --- p.25 / Chapter 2.3.2.3 --- Extraction of the GH Enriched DNA Fraction --- p.28 / Chapter 2.3.2.4 --- Cloning into Phage Vector Lambda GT11 --- p.29 / Chapter 2.3.2.5 --- Studies on the Enriched Genomic Library --- p.31 / Chapter 2.4 --- Result --- p.33 / Chapter 2.5 --- Discussion --- p.43 / Chapter Chapter 3 --- Screening --- p.46 / Chapter 3.1 --- Strategy --- p.46 / Chapter 3.2 --- Materials and Methods --- p.47 / Chapter 3.2.1 --- Materials --- p.47 / Chapter 3.2.2 --- Procedure --- p.48 / Chapter 3.2.2.1 --- Primary Screening --- p.48 / Chapter 3.2.2.2 --- Purificaiton of Positive Signal --- p.49 / Chapter 3.2.2.3 --- DNA Extraction from Positive Clone --- p.49 / Chapter 3.3 --- Result --- p.50 / Chapter 3.4 --- Discussion --- p.58 / Chapter Chapter 4 --- Studies on the Positive Clone --- p.59 / Chapter 4.1 --- Introduction --- p.59 / Chapter 4.2 --- Materials and Methods --- p.60 / Chapter 4.2.1 --- Materials --- p.60 / Chapter 4.2.2 --- Procedure --- p.62 / Chapter 4.2.2.1 --- Insert DNA Preparation --- p.62 / Chapter 4.2.2.2 --- Single Enzyme Digestion --- p.62 / Chapter 4.2.2.3 --- Double Enzyme Digestion --- p.62 / Chapter 4.2.2.4 --- Preparation of Position Specific Probes --- p.62 / Chapter 4.2.2.5 --- Hybridization with Position Specific Probes --- p.64 / Chapter 4.2.2.6 --- Preparation of Competent Cells --- p.65 / Chapter 4.2.2.7 --- Subcloning into pUC18 --- p.65 / Chapter 4.2.2.8 --- Plasmid Preparation --- p.66 / Chapter 4.2.2.9 --- Sequencing --- p.67 / Chapter 4.3 --- Result --- p.69 / Chapter 4.4 --- Discussion --- p.78 / Chapter Chapter 5 --- General Discussion --- p.80 / Appendix A --- p.83 / Appendix B --- p.93 / References --- p.100

Lai-Han Leung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 120-129). / Abstracts in English and Chinese. / Members of Thesis Advisory Committee --- p.i / Acknowledgments --- p.ii / Abstract --- p.iii / Abbreviations --- p.viii / Table of contents --- p.ix / Chapter Ch 1 --- General Introduction / Chapter 1.1 --- Growth hormone --- p.1 / Chapter 1.1.1 --- Physiology of GH --- p.1 / Chapter 1.1.2 --- Biological Functions of growth hormone --- p.2 / Chapter 1.1.3 --- Structure of growth hormone --- p.3 / Chapter 1.1.4 --- The physiological activities of growth hormone --- p.5 / Chapter 1.1.5 --- The importance of increasing grass carp production yield --- p.6 / Chapter 1.1.6 --- Application of recombinant growth hormone --- p.7 / Chapter 1.1.7 --- Application of recombinant grass carp GH --- p.8 / Chapter 1.2 --- Administration of GH to fish --- p.9 / Chapter 1.2.1 --- Methods in the administration of growth hormone to fish --- p.9 / Chapter 1.3 --- Different approaches in the expression of GH --- p.12 / Chapter 1.3.1 --- Bacterial expression system --- p.12 / Chapter 1.3.2 --- Baculovirus and yeast expression system --- p.14 / Chapter 1.4 --- Expression of grass carp growth hormone in yeast (Saccharomyces cerevisiae & Pichia Pastoris) --- p.14 / Chapter 1.4.1 --- Saccharomyces cerevisiae is widely used in the past --- p.14 / Chapter 1.4.2 --- Pichia pastoris is an alternative approach to overcome drawbacks of Saccharomyces cerevisiae --- p.16 / Chapter 1.5 --- Methods for increasing expression level of a cloned gene --- p.20 / Chapter 1.6 --- Purpose of present study --- p.22 / Chapter Ch 2 --- Materials and Method / Chapter 2.1 --- Bacterial strains --- p.23 / Chapter 2.2 --- Yeast strains --- p.23 / Chapter 2.3 --- Plasmids --- p.23 / Chapter 2.4 --- Bacterial culture Media and Solutions --- p.23 / Chapter 2.5 --- Antibiotic Solutions --- p.24 / Chapter 2.6 --- Restriction endonucleases and other enzymes --- p.24 / Chapter 2.7 --- Culture stock solutions --- p.24 / Chapter 2.8 --- SDS-PAGE and Western blot reagents --- p.28 / Chapter 2.9 --- DNA agarose gel --- p.30 / Chapter 2.10 --- General Techniques --- p.32 / Chapter 2.11 --- Construction of recombinant yeast strain --- p.40 / Chapter 2.12 --- Expression of the recombinant protein --- p.42 / Chapter 2.13 --- Purification of the protein --- p.44 / Chapter 2.14 --- Characterization of the protein --- p.46 / Chapter Ch3 --- Results / Chapter 3.1 --- Construction of Recombinant Plasmids --- p.50 / Chapter 3.2 --- Sequencing of the recombinants --- p.61 / Chapter 3.3 --- Linearization and electroporation of recombinant plasmids --- p.69 / Chapter 3.4 --- Optimization of electroporation condition of producing recombinant yeast cells --- p.70 / Chapter 3.5 --- Screening and selection of AOX characteristics (By MM plates) --- p.71 / Chapter 3.6 --- Screening and selection of AOX characteristics (By PCR) --- p.72 / Chapter 3.7 --- Selection of high expression clones --- p.77 / Chapter 3.8 --- Time course expression studies --- p.82 / Chapter 3.9 --- Batch fermentation --- p.89 / Chapter 3.10 --- Purification of r-gcGH --- p.92 / Chapter 3.11 --- Characterization of r-gcGH --- p.100 / Chapter 3.11.1 --- Immunological property of r-gcGH --- p.100 / Chapter 3.11.2 --- Biological activity of r-gcGH --- p.101 / Chapter Ch4 --- Discussion / Chapter 4.1 --- Evaluation of expression profile --- p.108 / Chapter 4.1.1 --- Expression yield --- p.108 / Chapter 4.1.2 --- Efficiency in the cleavage of signal peptide --- p.111 / Chapter 4.2 --- Evaluation of the biological activity of r-gcGH --- p.113 / Chapter 4.3 --- Evaluation of the post-translational modification of r-gcGH --- p.115 / Chapter 4.4 --- Further studies --- p.116 / Reference --- p.120

Thesis (M. Phil.)--University of Hong Kong, 1998. / Includes bibliographical references (leaves 59-75).

published_or_final_version / abstract / Biological Sciences / Doctoral / Doctor of Philosophy

by Agnes Pui-Yee Chan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1992. / Includes bibliographical references (leaves 162-177). / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.iv / ABBREVIATIONS --- p.vii / Chapter CHAPTER 1 --- INTRODUCTION / Chapter 1.1 --- Physiology of growth --- p.3 / Chapter 1.2 --- The anterior pituitary --- p.4 / Chapter 1.3 --- Chemistry of GH and the GH gene family --- p.7 / Chapter 1.4 --- Biochemical effects and mode of action of GH --- p.8 / Chapter 1.5 --- Control of GH at cellular level --- p.10 / Chapter 1.6 --- Control of GH gene expression at molecular level / Chapter 1.6.1 --- Introduction --- p.11 / Chapter 1.6.2 --- Tissue-specific expression of GH gene / Chapter 1.6.2.1 --- Tissue-specific transcription factors of pituitary cells --- p.20 / Chapter 1.6.2.2 --- Non-tissue specific transcription factors of pituitary cells --- p.27 / Chapter 1.6.2.3 --- Negatively-acting transcription factors of non-pituitary cells --- p.34 / Chapter 1.6.2.4 --- Theory for tissue-specific GH gene activation --- p.39 / Chapter 1.7 --- Characteristic of growth in fish --- p.40 / Chapter 1.8 --- Objectives of the present study --- p.42 / Chapter CHAPTER 2 --- MATERIALS AND METHODS / Chapter 2.1 --- General techniques / Chapter 2.1.1 --- Preparation of DNA / Chapter 2.1.1.1 --- Minipreparation of DNA --- p.46 / Chapter 2.1.1.2 --- Preparation of DNA using Qiagen column --- p.47 / Chapter 2.1.1.3 --- Preparation of phage DNA --- p.48 / Chapter 2.1.2 --- Elution of DNA from agarose gel --- p.51 / Chapter 2.1.3 --- Preparation of competence cells and transformation --- p.52 / Chapter 2.1.4 --- Ligation of DNA fragments --- p.53 / Chapter 2.1.5 --- Cell feeding and subculturing --- p.54 / Chapter 2.2 --- Special techniques / Chapter 2.2.1 --- DNA sequencing --- p.56 / Chapter 2.2.2 --- Polymerase chain reaction (PCR) --- p.67 / Chapter 2.2.3 --- Direct sequencing of PCR products --- p.72 / Chapter 2.2.4 --- Nested-deletion --- p.75 / Chapter 2.2.5 --- DNA transfection --- p.81 / Chapter 2.2.6 --- CAT assay --- p.86 / Chapter CHAPTER 3 --- RESULTS / Chapter 3.1 --- Sequencing of the grass carp GH gene / Chapter 3.1.1 --- Introduction --- p.93 / Chapter 3.1.2 --- Sequencing strategy --- p.94 / Chapter 3.2 --- Sequence analysis of the grass carp GH gene --- p.108 / Chapter 3.3 --- Functional analysis of the grass carp GH gene --- p.115 / Chapter CHAPTER 4 --- DISCUSSIONS / Chapter 4.1 --- DNA sequence comparison between grass carp GH gene and other organisms --- p.137 / Chapter 4.2 --- Amino acid comparisons between grass carp GH and other organisms --- p.143 / Chapter 4.3 --- Tissue-specific expression of GH gene / Activation of transcription --- p.154 / Repression of transcription --- p.155 / Chapter 4.4 --- Electroporation of zebrafish eggs --- p.157 / Chapter 4.5 --- Further studies --- p.160 / REFERENCES --- p.162 / APPENDIX --- p.178

Poon, Chi-to, Geoffrey. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 115-125). / Acknowledgements --- p.I / Abbreviations --- p.II / Abstract --- p.III / Table of content --- p.IV / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Importance of growth enhancement in aquaculture --- p.1 / Chapter 1.2 --- Physiological effect of growth hormone --- p.1 / Chapter 1.3 --- Progress in teleost growth hormone research --- p.3 / Chapter 1.4 --- Grass carp and its aquaculture --- p.5 / Chapter 1.5 --- Route of administration of growth hormone --- p.8 / Chapter 1.6 --- Nomenclature of baculovirus --- p.9 / Chapter 1.7 --- Biology of baculovirus --- p.10 / Chapter 1.8 --- Control of gene expression of virus-infected cells --- p.13 / Chapter 1.9 --- Theme of the thesis --- p.14 / Chapter Chapter 2 --- Materials and Methods --- p.18 / Chapter 2.1 --- Synthesis and purification of primers --- p.18 / Chapter 2.2 --- Modification of gcGH cDNA by polymerase chain reaction (PCR) --- p.20 / Chapter 2.3 --- TA cloning of PCR product --- p.20 / Chapter 2.4 --- Purification ofDNA fragment from agarose gel by GENECLEAN´ёØ --- p.21 / Chapter 2.5 --- Recovery of low molecular weight DNA fragment from agarose gel --- p.22 / Chapter 2.6 --- Small scale preparation of plasmid DNA --- p.23 / Chapter 2.7 --- Large scale plasmid preparation by QIAGEN´ёØ --- p.24 / Chapter 2.8 --- Preparation of competent Escherichia coli JM109 for transformation --- p.25 / Chapter 2.9 --- Transformation of plasmid into competent Escherichi coli JM109 --- p.26 / Chapter 2.10 --- Cell culture of BmN cell line --- p.26 / Chapter 2.10.1 --- Preparation of TC-100 insect medium --- p.27 / Chapter 2.10.2 --- Preparation of Grace's medium --- p.27 / Chapter 2.11 --- Extraction of wild-type Bombyx mori nuclear polyhedrosis virus DNA --- p.28 / Chapter 2.12 --- Transfection of BmN cells with Bombyx mori nuclear polyhedrosis virus DNA by DOTAP´ёØ --- p.28 / Chapter 2.13 --- Agarose plaque assay --- p.29 / Chapter 2.14 --- Lifting of vius plaque onto nitrocellulose filter paper --- p.30 / Chapter 2.15 --- Synthesis of radiolabelled DNA probe --- p.31 / Chapter 2.16 --- Pre-hybridization and hybridization of recombinant virus DNA on nitrocellulose paper --- p.31 / Chapter 2.17 --- Purification of recombinant virus by dot-blot manifold --- p.33 / Chapter 2.18 --- Preparation of cell lysate from virus-infected BmN cells --- p.33 / Chapter 2.19 --- Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.34 / Chapter 2.19.1 --- Staining of the gel by Coomassie blue method --- p.35 / Chapter 2.19.2 --- Staining of the gel by silver staining method --- p.35 / Chapter 2.20 --- Determination of protein concentration by Bradford's method --- p.36 / Chapter 2.21 --- Determination of total protein concentration by Folin-Lowry method --- p.36 / Chapter 2.22 --- Detection of grass carp growth hormone by Western blotting --- p.37 / Chapter 2.23 --- Preparation of native recombinant grass carp growth hormone for iodination --- p.38 / Chapter 2.24 --- Iodination of recombinat grass carp growth hormone by IODO-GEN´ёØ --- p.38 / Chapter 2.25 --- Purification of radiolabelled recombinant grass carp growth hormone --- p.39 / Chapter 2.26 --- Radioimmunoassay (RIA) for detection of recombinant grass carp growth hormone --- p.40 / Chapter 2.27 --- Ammonium sulphate precipitation --- p.41 / Chapter Chapter 3 --- Vector Construction --- p.42 / Chapter 3.1 --- Components of parent vector pBM030 --- p.42 / Chapter 3.2 --- Construction of pBM-EE --- p.44 / Chapter 3.3 --- Constrcution of pBM-EX --- p.47 / Chapter Chapter 4 --- Results --- p.51 / Chapter 4.1 --- Construction and purfication of recombinant baculovirus --- p.51 / Chapter 4.2 --- Expression of recombinant grass carp growth hormone in BmN cells --- p.55 / Chapter 4.3 --- Expression of recombinant grass carp growth hormone in Bombyx mori larva --- p.62 / Chapter 4.4 --- Putative physical characteristics of the recombinant grass carp growth hormone --- p.67 / Chapter 4.5 --- Purification of the grass carp growth hormone in Bombyx mori larva --- p.69 / Chapter 4.5.1 --- Ammonium sulphate precipitation --- p.69 / Chapter 4.5.2 --- Gel filtration --- p.72 / Chapter 4.5.3 --- Hydrophobic interaction chromatography --- p.75 / Chapter 4.5.4 --- Anion exchange chromatography --- p.78 / Chapter 4.5.5 --- Reverse phase chromatography --- p.90 / Chapter Chapter 5 --- Discussions --- p.99 / Chapter 5.1 --- Merits of baculovirus expression system against other expression systems --- p.99 / Chapter 5.2 --- Basic design of the recombinant baculovirus transfer vector --- p.100 / Chapter 5.3 --- Potential for Mutation of the Baculovirus during Homologous Recombination --- p.101 / Chapter 5.4 --- Cleavage of Signal Peptide from the Expressed Protein --- p.103 / Chapter 5.5 --- Difference in recombinant gcGH expression levelin EE4-7 and EX3-16 --- p.103 / Chapter 5.6 --- Purification of recombinant gcGH protein --- p.106 / Chapter 5.6.1 --- Chromatographic behaviour of recombinant gcGH in Q-Sepharose column --- p.106 / Chapter 5.6.2 --- Problem of aggregation of recombinant gcGH --- p.107 / Chapter 5.6.3 --- Solvent system used in recombinant gcGH purification --- p.108 / Chapter 5.6.4 --- Protein denaturating effect of the solvent system --- p.109 / Chapter 5.6.5 --- Protein yield --- p.110 / Chapter 5.7 --- Problems and accuracy of radioimmunoassay --- p.110 / Chapter Chapter 6 --- Further study --- p.113 / Chapter Chapter 7 --- References --- p.115 / Appendix I --- p.126 / Appendix II: Construction of the Supervector --- p.127