Chan Hiu-ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 97-112). / Abstracts in English and Chinese. / Thesis Committee --- p.i / 摘要 --- p.ii / Abstract --- p.iii / Acknowledgements --- p.v / Abbreviations --- p.vi / Table of Contents --- p.vii / List of figures --- p.xi / List of tables --- p.xiii / Chapter Chapter 1 --- Literature Review --- p.1 / Chapter 1.1 --- Nitrogen assimilation in plants --- p.1 / Chapter 1.2 --- Importance of asparagine in plants --- p.5 / Chapter 1.3 --- Enzymatic reaction of asparagine synthetase (AS) --- p.8 / Chapter 1.4 --- Asparagine synthetase of non-plant organisms --- p.10 / Chapter 1.5 --- Biochemistry background of plant asparagine synthetases --- p.12 / Chapter 1.6 --- Molecular studies of asparagine synthetase genes in plants --- p.15 / Chapter 1.7 --- Arabidopsis thaliana as a model plant --- p.24 / Chapter 1.8 --- ASN studies in Arabidopsis thaliana --- p.24 / Chapter 1.9 --- Hypothesis --- p.27 / Chapter Chapter 2 --- Materials and Methods --- p.29 / Chapter 2.1 --- Chemicals --- p.29 / Chapter 2.2 --- Plant materials and growth conditions --- p.29 / Chapter 2.2.1 --- Surface sterilization of Arabidopsis seeds --- p.29 / Chapter 2.2.2 --- "Growth conditions of Arabidopsis seeds for total RNA extraction, enzyme assay, chlorophyll content measurement and dry weight measurement" --- p.30 / Chapter 2.3 --- Agrobacterium mediated transformation via vacuum infiltration method --- p.30 / Chapter 2.3.1 --- Principles --- p.30 / Chapter 2.3.2 --- Plant materials and bacterial strains of Agrobacterium mediated transformation --- p.31 / Chapter 2.3.2.1 --- Plant materials --- p.31 / Chapter 2.3.2.2 --- Gene constructs --- p.31 / Chapter 2.3.2.2 --- Bacterial strains --- p.32 / Chapter 2.3.3 --- Agrobacterium mediated transformation via vacuum infilitration --- p.32 / Chapter 2.4 --- Screening of transformants --- p.33 / Chapter 2.5 --- DNA and RNA manipulation --- p.34 / Chapter 2.5.1 --- DNA extraction and quantitation --- p.34 / Chapter 2.5.2 --- PCR amplification and detection of transgenes --- p.36 / Chapter 2.5.2.1 --- PCR amplification and detection of transgenes --- p.36 / Chapter 2.5.2.2 --- Primer sequence --- p.37 / Chapter 2.6 --- RNA analysis of transormants --- p.38 / Chapter 2.6.1 --- General introduction --- p.38 / Chapter 2.6.2 --- RNA extraction --- p.39 / Chapter 2.6.3 --- Making single-strand DIG PCR probes --- p.40 / Chapter 2.6.4 --- Quantitation of single-strand DIG-labeled probes --- p.42 / Chapter 2.7 --- Northern blot analysis --- p.42 / Chapter 2.7.1 --- Detection --- p.43 / Chapter 2.7.2 --- Film development --- p.43 / Chapter 2.8 --- "Amino acid, protein, dry weight and total nitrogen analysis" --- p.43 / Chapter 2.8.1 --- Extraction of free amino acids --- p.43 / Chapter 2.8.2 --- Protein assay --- p.44 / Chapter 2.8.3 --- Determination of nitrogen and carbon content in seeds --- p.45 / Chapter 2.8.4 --- Dry weight measurement --- p.45 / Chapter 2.8.5 --- Seed storage protein analyses --- p.45 / Chapter 2.8.6 --- Detection of chlorophyll content --- p.46 / Chapter 2.9 --- Asparagine synthetase activity analysis --- p.46 / Chapter 2.9.1 --- Crude extracts preparation --- p.46 / Chapter 2.9.2 --- AS enzyme assay --- p.47 / Chapter 2.9.3 --- Asparagine content measurement --- p.47 / Chapter 2.10 --- In situ hybridization --- p.48 / Chapter 2.10.1 --- Making cRNA probe --- p.48 / Chapter 2.10.2 --- In situ hybridization --- p.48 / Chapter Chapter 3 --- Results --- p.50 / Chapter 3.1 --- Construction of ASN1 overexpressing lines --- p.50 / Chapter 3.2 --- Changes in nitrogen status during vegetative growth of ASN1 overexpressing lines --- p.53 / Chapter 3.3 --- Changes in nitrogen status during seed development of ASN1 overexpressing lines --- p.55 / Chapter Chapter 4 --- Discussion --- p.76 / Chapter Chapter 5 --- Conclusion --- p.85 / Chapter Chapter 6 --- Perspective --- p.86 / Appendix --- p.87 / References --- p.97
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323219 |
| Date | January 2000 |
| Contributors | Chan, Hiu-ki., 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, xiii, 112 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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