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
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324439 |
| Date | January 2003 |
| Contributors | Leung, Lai-Han., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, x, 129 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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