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Holocentromeres and the centromeric histone H3 proteins.

Cheung, Wai Kuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 66-76). / Abstracts in English and Chinese. / List of Figures --- p.v / List of Tables --- p.vi / List of Abbreviations --- p.vii / Acknowledgements --- p.ix / Abstract --- p.xi / 摘要 --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Centromere and Kinetochore --- p.1 / Chapter 1.2 --- The Kinetochore Subunits: Centromeric Nucleosomes --- p.2 / Chapter 1.3 --- CenH3: The Centromere Specific Histone --- p.5 / Chapter 1.4 --- The Centromeric DNA: Tandem Repeats and Retrotransposons --- p.8 / Chapter 1.5 --- The Genetic and Epigenetic Nature of the Centromeres --- p.9 / Chapter 1.6 --- Point Centromeres and Regional Centromeres --- p.10 / Chapter 1.7 --- Holocentric Chromosomes --- p.11 / Chapter 1.8 --- Hypothesis --- p.13 / Chapter Chapter 2 --- Materials and methods --- p.15 / Chapter 2.1 --- Chemicals --- p.15 / Chapter 2.2 --- Bacterial strains in routine cloning --- p.15 / Chapter 2.3 --- Plant materials in cloning and transformation --- p.15 / Chapter 2.4 --- Construction of LnCENH3-GFP and CeHCP3-DsRED chimeric gene cassettes for rice transformation --- p.15 / Chapter 2.5 --- Cloning of CENH3 gene of Luzula spp --- p.22 / Chapter 2.6 --- Construction of full length OsCENH3 RNAi and 150bp OsCENH3 RNAi constructs for rice transformation --- p.22 / Chapter 2.7 --- Agrobacterium-mediated transformation of rice (Oryza sativa L.japonica. cv. Nipponbare) --- p.24 / Chapter 2.8 --- Gene gun transformation of rice (Oryza sativa L.japonica. cv. Nipponbare) by Biolistic PDS-1000/He´ёØ System (Bio-rad) --- p.26 / Chapter 2.9 --- Detection of transgenes expression --- p.28 / Chapter 2.10 --- Nuclear protein extraction --- p.29 / Chapter 2.11 --- Protein-DNA Binding Assay --- p.30 / Chapter 2.12 --- Protein precipitation by methanol-chloroform --- p.32 / Chapter 2.13 --- Western blot analysis of proteins from Protein-DNA binding assay --- p.33 / Chapter 2.14 --- Tubulin immunolocalization of root tips --- p.33 / Chapter 2.15 --- Bioinformatics analysis --- p.34 / Chapter Chapter 3 --- Results --- p.36 / Chapter 3.1 --- Plant transformation vectors construction --- p.36 / Chapter 3.2 --- Rice transformation --- p.38 / Chapter 3.3 --- Transgenic plants screening --- p.39 / Chapter 3.4 --- Analysis of the codon usages of CeHCP-3 gene in C. elegans and O. sativa --- p.42 / Chapter 3.5 --- In vitro Protein-DNA binding assays --- p.44 / Chapter 3.6 --- Subcellular localization study of LnCENH3 in rice --- p.46 / Chapter 3.7 --- Chromosome morphology of the transgenic rice expression LnCENH3 --- p.47 / Chapter 3.8 --- Tubulin immunolocalization of LnCENH3-GFP transgenic rice --- p.49 / Chapter 3.9 --- Cloning of CENH3s in Luzula genus --- p.51 / Chapter 3.10 --- Bioinformatics analysis of Luzula CENH3s --- p.53 / Chapter Chapter 4 --- Discussion --- p.57 / Chapter 4.1 --- Expression of fusion proteins in rice --- p.57 / Chapter 4.2 --- Incorporation of LnCENH3-GFP in nucleosomes --- p.57 / Chapter 4.3 --- Expression pattern of LnCENH3-GFP in rice --- p.58 / Chapter 4.4 --- Formation of additional kinetochores on transgenic rice chromosome --- p.60 / Chapter 4.5 --- Deletion of 8 amino acids in LeCENH3 --- p.62 / Chapter Chapter 5 --- Conclusion --- p.65 / References --- p.66 / Chapter 5.1 --- Appendix --- p.77

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_327362
Date January 2011
ContributorsCheung, Wai Kuen., Chinese University of Hong Kong Graduate School. Division of Life Sciences.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xiii, 79 leaves : ill. (chiefly col.) ; 30 cm.
RightsUse 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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