by Chan Wah Lun. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 252-260). / Acknowledgments --- p.i / Abstract --- p.ii / Contents --- p.vi / Naming of TCS mutants and modified TCS protein --- p.x / Abbreviations --- p.xi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Physical and chemical properties of Trichosanthin --- p.1 / Chapter 1.2 --- Biological activities of Trichosanthin --- p.3 / Chapter 1.3 --- Renal tubular reabsorption and nephrotoxicity of Trichosanthin --- p.10 / Chapter 1.4 --- Objective and strategies of study --- p.11 / Chapter Chapter 2 --- Materials and methods --- p.19 / Chapter 2.1 --- General Techniques --- p.19 / Chapter 2.2 --- Site directed mutagenesis of Trichosanthin --- p.21 / Chapter 2.3 --- DNA sequencing --- p.37 / Chapter 2.4 --- Overexpression of modified Trichosanthin in E. coli --- p.42 / Chapter 2.5 --- Purification of modified Trichosanthin --- p.43 / Chapter 2.6 --- Breaking of Disulphide bridge between modified TCS --- p.44 / Chapter 2.7 --- Coupling of DX T40 to modified Trichosanthin --- p.44 / Chapter 2.8 --- Biological activities of modified Trichosanthin and Dextran-modified trichosantin conjugates --- p.46 / Chapter 2.9 --- Immunogenicity of modified Trichosanthin and Dextran-trichosanthin conjugates --- p.50 / Chapter 2.10 --- Nephrotoxicity of Trichosanthin and Dextran-trichosanthin conjugates --- p.53 / Chapter Chapter 3 --- Construction of TCS mutants --- p.61 / Chapter 3.1 --- Introduction --- p.61 / Chapter 3.2 --- Method --- p.61 / Chapter 3.3 --- Results --- p.62 / Chapter 3.3.1 --- Construction of K173C mutant --- p.62 / Chapter 3.3.2 --- Construction of R29C mutant --- p.64 / Chapter 3.3.3 --- Construction of K173C R29C double mutant --- p.65 / Chapter 3.4 --- Discussion --- p.66 / Chapter Chapter 4 --- "Expression,Purification and Ribosome- inactivating activities of Modified Trichosanthin proteins" --- p.87 / Chapter 4.1 --- Introduction --- p.87 / Chapter 4.2 --- Method --- p.87 / Chapter 4.3 --- Results --- p.88 / Chapter 4.3.1 --- "Expression, purification and ribosome-inactivating activity of K173C" --- p.88 / Chapter 4.3.2 --- "Expression ,purification and ribosome-inactivating activity of R29C" --- p.89 / Chapter 4.3.3 --- "Expression, purification and ribosome-inactivating activity of K173C R29C" --- p.90 / Chapter 4.4 --- Discussion --- p.91 / Chapter Chapter 5 --- Coupling of Dextran T40 to modified Trichosanthin --- p.108 / Chapter 5.1 --- Introduction --- p.108 / Chapter 5.2 --- Method --- p.109 / Chapter 5.3 --- Results --- p.109 / Chapter 5.3.1 --- Coupling of R29C --- p.109 / Chapter 5.3.2 --- Coupling of K173C --- p.111 / Chapter 5.3.3 --- Coupling of R29CK173C --- p.111 / Chapter 5.4 --- Discussion --- p.111 / Chapter Chapter 6 --- Biological Activities of modified Trichosanthin and Dextran-modified trichosanthin conjugates --- p.128 / Chapter 6.1 --- Introduction --- p.128 / Chapter 6.2 --- Method --- p.128 / Chapter 6.3 --- Results --- p.130 / Chapter 6.3.1 --- In vivo Biological activity- Mid-term abortifacient activity --- p.130 / Chapter 6.3.2 --- In vitro biological activities / Chapter 6.3.2a --- Ribosome-inactivating activity --- p.131 / Chapter 6.3.2b --- Anti-tumour activity --- p.132 / Chapter 6.4 --- Discussion --- p.133 / Chapter Chapter 7 --- Immunogenicity of Dextran-modified trichosanthin conjugates --- p.156 / Chapter 7.1 --- Introduction --- p.156 / Chapter 7.2 --- Method --- p.157 / Chapter 7.3 --- Results / Chapter 7.3.1 --- Immunogenicity without denaturation of protein --- p.158 / Chapter 7.3.2 --- Immunogenicity with denaturation of protein --- p.161 / Chapter 7.4 --- Discussion --- p.162 / Chapter Chapter 8 --- Nephrotoxicity of Trichosanthin and Dextran-Trichosanthin conjugates --- p.199 / Chapter 8.1 --- Introduction --- p.200 / Chapter 8.2 --- Method --- p.202 / Chapter 8.3 --- Results --- p.202 / Chapter 8.3.1 --- Functional study on nephrotoxicity of Trichosanthin --- p.202 / Chapter 8.3.2 --- Morphological study on the nephrotoxicity of Trichosanthin --- p.203 / Chapter 8.3.3 --- The effect of coupling of Dextran T40 on the nephrotoxicity of Trichosanthin --- p.206 / Chapter 8.4 --- Discussion --- p.207 / Chapter Chapter 9 --- General Discussion --- p.244 / References --- p.252
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318343 |
| Date | January 1995 |
| Contributors | Chan, Wah Lun., Chinese University of Hong Kong Graduate School. Division of Basic Medical Sciences. |
| Publisher | Chinese University of Hong Kong |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xiv, 260 leaves : ill. (some mounted 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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