Wong Wai Mei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 175-192). / Abstracts in English and Chinese. / Declaration --- p.ii / Acknowledgements --- p.iii / Abstract --- p.iv / Abbreviation --- p.vi / Table of Contents --- p.vii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Section I --- The Making of Genetically Modified Organisms --- p.2 / Chapter 1.1 --- Conventional breeding in agriculture --- p.2 / Chapter 1.2 --- What is genetic engineering? --- p.4 / Chapter 1.3 --- Plant transformation --- p.5 / Chapter 1.3.1 --- Agrobacterium-mediated --- p.6 / Chapter 1.3.2 --- Direct gene transfer --- p.8 / Chapter 1.3.2.1 --- Microparticle bombardment --- p.8 / Chapter 1.3.2.2 --- Protoplasts --- p.9 / Chapter 1.3.3 --- Gene silencing --- p.10 / Chapter 1.4 --- Examples of genetically modified crops --- p.13 / Chapter 1.5 --- Foreign genes commonly found in transgenic plants --- p.14 / Chapter Section II --- Benefits and Environmental Concern of GMOs --- p.17 / Chapter 2.1 --- Mechanism of GMO --- p.17 / Chapter 2.1.1 --- Herbicide tolerant crops --- p.18 / Chapter 2.1.2 --- Insect resistant crops --- p.19 / Chapter 2.1.3 --- Delayed ripening crops --- p.20 / Chapter 2.1.4 --- Virus resistant crops --- p.20 / Chapter 2.2 --- Benefits of GMOs --- p.21 / Chapter 2.3 --- Impact of GM foods to human health and the environment --- p.22 / Chapter 2.3.1 --- Human health --- p.22 / Chapter 2.3.1.1 --- GM potatoes --- p.23 / Chapter 2.3.1.2 --- CaMV risks? --- p.24 / Chapter 2.3.1.3 --- Food allergy --- p.25 / Chapter 2.3.2 --- Environmental concerns --- p.26 / Chapter 2.3.2.1 --- Horizontal gene transfer --- p.27 / Chapter 2.3.2.1.1 --- Selectable marker genes --- p.27 / Chapter 2.3.2.1.2 --- Herbicide resistant genes --- p.29 / Chapter 2.3.2.1.3 --- Insect resistant genes --- p.29 / Chapter 2.3.2.2 --- Ecology --- p.30 / Chapter 2.3.2.2.1 --- Monarch butterfly --- p.30 / Chapter Section III --- Future developments of GMO --- p.32 / Chapter 3.1 --- Designer Food and engineered plants --- p.32 / Chapter 3.1.1 --- Insect resistance --- p.33 / Chapter 3.1.2 --- Viral resistance --- p.33 / Chapter 3.1.3 --- Fungal resistance --- p.34 / Chapter 3.1.4 --- Nutritional quality --- p.34 / Chapter 3.1.5 --- Modifications of oil composition --- p.35 / Chapter 3.1.6 --- Medical applications --- p.37 / Chapter 3.1.7 --- Environmental applications --- p.40 / Chapter 3.1.7.1 --- Tolerance to high salinity and drought --- p.40 / Chapter 3.1.7.2 --- Tolerance to frost --- p.41 / Chapter 3.1.7.3 --- Bioremediation --- p.42 / Chapter 3.1.7.4 --- Biodegradable products --- p.43 / Chapter Section IV --- Regulation of GMO --- p.44 / Chapter 4.1 --- The question of labeling --- p.44 / Chapter 4.1.1 --- Moral and ethical issues --- p.44 / Chapter 4.1.2 --- Animal welfare --- p.45 / Chapter 4.2 --- International practice in GMO labeling --- p.46 / Chapter 4.2.1 --- United States of America --- p.46 / Chapter 4.2.2 --- Canada --- p.48 / Chapter 4.2.3 --- European Union --- p.49 / Chapter 4.2.4 --- Australia and New Zealand --- p.50 / Chapter 4.2.5 --- Japan --- p.51 / Chapter 4.2.6 --- Republic of Korea --- p.52 / Chapter 4.2.7 --- China --- p.53 / Chapter 4.2.8 --- Taiwan --- p.53 / Chapter 4.2.9 --- Hong Kong --- p.54 / Chapter Section V --- Uses of crops --- p.56 / Chapter 5.1 --- Uses of crops --- p.56 / Chapter 5.1.1 --- Soybean --- p.56 / Chapter 5.1.2 --- Corn --- p.57 / Chapter 5.1.3 --- Tomato --- p.58 / Chapter 5.1.4 --- Potato --- p.59 / Chapter 5.1.5 --- Rice --- p.60 / Chapter 5.1.6 --- Rapeseed --- p.61 / Chapter 5.1.7 --- Oil --- p.62 / Chapter 5.2 --- "Food additives, hormones and flavourings" --- p.63 / Chapter Chapter 2 --- Materials & Methods --- p.65 / Chapter 2.1 --- Materials --- p.66 / Chapter 2.1.1 --- Growth media & agar --- p.66 / Chapter 2.1.2 --- Reagents for agarose gel electrophoresis --- p.67 / Chapter 2.1.3 --- Reagents for preparation of competent cells --- p.67 / Chapter 2.1.4 --- Reagents for measurement of DNA concentration --- p.68 / Chapter 2.1.4.1 --- Measurement of DNA concentration by PicoGreen --- p.68 / Chapter 2.1.5 --- Reagents for Southern hybridization --- p.68 / Chapter 2.2 --- Methods --- p.70 / Chapter 2.2.1 --- Restriction endonuclease digestion --- p.70 / Chapter 2.2.2 --- Agarose gel electrophoresis of DNA --- p.70 / Chapter 2.2.3 --- DNA recovery from agarose gel --- p.71 / Chapter 2.2.3.1 --- QIAquick® gel extraction --- p.71 / Chapter 2.2.4 --- Ligation of purified DNA fragment into vector --- p.72 / Chapter 2.2.5 --- Transformation --- p.72 / Chapter 2.2.6 --- Rubidium chloride method for making competent cells --- p.12 / Chapter 2.2.7 --- Plasmid DNA preparation --- p.73 / Chapter 2.2.7.1 --- Concert Rapid Mini Prep --- p.73 / Chapter 2.2.7.2 --- QIAprep® Miniprep --- p.74 / Chapter 2.2.8 --- Extraction of plant genomic DNA --- p.75 / Chapter 2.2.8.1 --- Qiagen DNeasy´ёØ Plant Mini Kit --- p.75 / Chapter 2.2.9 --- Southern Hybridization --- p.75 / Chapter 2.2.9.1 --- Denaturation --- p.76 / Chapter 2.2.9.2 --- Blot transfer --- p.76 / Chapter 2.2.9.3 --- Pre-hybridization --- p.77 / Chapter 2.2.9.4 --- Synthesis of radiolabelled probe --- p.77 / Chapter 2.2.9.5 --- Hybridization of radiolabelled probe on filter --- p.77 / Chapter 2.2.9.6. --- Detection of hybridized probes --- p.78 / Chapter 2.2.10 --- Measurement of DNA concentration --- p.78 / Chapter 2.2.10.1 --- Determination of DNA on EtBr stained gel --- p.78 / Chapter 2.2.10.2 --- Determination of DNA by UV spectrophotometer --- p.78 / Chapter 2.2.10.3 --- Determination of DNA by PicoGreen --- p.79 / Chapter 2.2.11 --- DNA sequencing --- p.80 / Chapter 2.2.11.1 --- Automated sequencing by ABI Prism 377 --- p.80 / Chapter Chapter 3 --- PCR Diagnostics --- p.81 / Chapter 3.1 --- Applications of PCR to processed foods --- p.82 / Chapter 3.1.1 --- DNA quality --- p.82 / Chapter 3.1.2 --- PCR & Multiplex PCR --- p.83 / Chapter 3.1.3 --- Choice of primers --- p.84 / Chapter 3.1.4 --- Inhibitors --- p.84 / Chapter 3.2 --- Materials & Methods --- p.85 / Chapter 3.2.1 --- Selection of primers --- p.85 / Chapter 3.2.2 --- Amplification of target sequences --- p.86 / Chapter 3.2.3 --- Multiple amplification of target sequences --- p.87 / Chapter 3.3 --- Results --- p.88 / Chapter 3.4 --- Discussion --- p.93 / Chapter Chapter 4 --- Quality Control in GMO detection --- p.95 / Chapter 4.1 --- Standardization of pre- and post- PCR analysis --- p.96 / Chapter 4.1.1 --- General guidelines --- p.96 / Chapter 4.1.2 --- UV irradiation --- p.97 / Chapter 4.1.3 --- Inactivation protocols --- p.93 / Chapter 4.1.4 --- Positive and negative controls --- p.99 / Chapter 4.1.5 --- PCR verification --- p.99 / Chapter 4.1.6 --- Equipment decontamination --- p.100 / Chapter 4.2 --- Materials & Methods --- p.101 / Chapter 4.2.1 --- Selection of primers for external control --- p.101 / Chapter 4.2.2 --- Development of the external control --- p.101 / Chapter 4.2.3 --- Selection of primers for internal control --- p.103 / Chapter 4.3 --- Results --- p.104 / Chapter 4.4 --- Discussion --- p.107 / Chapter Chapter 5 --- DNA extraction from food samples --- p.110 / Chapter 5.1 --- Introduction --- p.111 / Chapter 5.2 --- Reagents and Buffers for DNA extraction from food samples --- p.112 / Chapter 5.2.1 --- Cetyltrimethylammonium bromide (CTAB) extraction method --- p.112 / Chapter 5.2.2 --- Organic-based extraction method --- p.113 / Chapter 5.2.3 --- Potassium acetate/sodium dodecyl sulphate precipitation method --- p.113 / Chapter 5.2.4 --- Hexane-based extraction method --- p.114 / Chapter 5.3 --- Weight and names of samples --- p.115 / Chapter 5.4 --- DNA extraction methods --- p.115 / Chapter 5.4.1 --- CTAB extraction method --- p.115 / Chapter 5.4.2 --- Qiagen DNeasy´ёØ plant mini kit --- p.116 / Chapter 5.4.3 --- Promega Wizard® genomic DNA purification --- p.116 / Chapter 5.4.4 --- Promega Wizard® Magnetic DNA purification system --- p.117 / Chapter 5.4.5 --- Promega Wizard® DNA Clean-Up system --- p.118 / Chapter 5.4.6 --- Qiagen QIAshreddrer´ёØ and QIAamp spin column --- p.119 / Chapter 5.4.7 --- Chelex-based extraction method --- p.119 / Chapter 5.4.8 --- Organic-based extraction method --- p.120 / Chapter 5.4.9 --- Nucleon PhytoPure extraction and purification method --- p.120 / Chapter 5.4.10 --- Potassium acetate/SDS precipitation method --- p.121 / Chapter 5.4.11 --- Hexane-based extraction method --- p.122 / Chapter 5.5 --- Results --- p.123 / Chapter 5.5.1 --- Comparison of eleven extraction methods --- p.123 / Chapter 5.5.2 --- Comparison of DNA extraction on selected methods --- p.125 / Chapter 5.6 --- Discussion --- p.132 / Chapter Chapter 6 --- Quantitative Analysis --- p.136 / Chapter 6.1 --- Introduction --- p.137 / Chapter 6.1.1 --- Chemistry of quantitative PCR --- p.138 / Chapter 6.1.2 --- PCR system --- p.140 / Chapter 6.2 --- Materials & Methods --- p.142 / Chapter 6.2.1 --- Design of primers and probes --- p.142 / Chapter 6.2.2 --- Methods --- p.145 / Chapter 6.3 --- Results --- p.146 / Chapter 6.3.1 --- Selection of primer/probe --- p.146 / Chapter 6.3.2 --- Primer optimization --- p.149 / Chapter 6.3.3 --- Quantitative analysis of real samples --- p.158 / Chapter 6.4 --- Discussion --- p.152 / Chapter Chapter 7 --- Conclusion --- p.168 / References --- p.175 / Appendix --- p.193
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323476 |
| Date | January 2001 |
| Contributors | Wong, Wai Mei., 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, xii, 193 leaves : ill. (some col., some mounted) ; 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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