by Yin Cheung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 146-164). / Acknowledgments --- p.i / Abstract --- p.ii / Abbreviations --- p.iv / Chapter CHAPTER 1 --- General Introduction --- p.1 / Chapter 1.1 --- Drosophila as a model for studying development --- p.1 / Chapter 1.2 --- The formation of the body axes of Drosophila --- p.3 / Chapter 1.2.1 --- Oogenesis --- p.5 / Chapter 1.2.2 --- Embryogenesis --- p.15 / Chapter 1.2.3 --- Segmentation --- p.16 / Chapter 1.2.4 --- Life cycle --- p.20 / Chapter 1.3 --- Egg-polarity genes are essential for development --- p.22 / Chapter 1.4 --- Maternal gene bicoid is required for formation of anterior structures in the embryo --- p.24 / Chapter 1.4.1 --- Phenotypes of bicoid mutant --- p.24 / Chapter 1.4.2 --- Transplantation experiment --- p.26 / Chapter 1.4.3 --- Establishment of an anterior to posterior bicoid protein gradient --- p.26 / Chapter 1.4.4 --- Localization step of bicoid mRNA --- p.27 / Chapter 1.4.5 --- Formation of bicoid protein gradient --- p.28 / Chapter 1.4.6 --- The bicoid protein gradient regulates the downstream zygotic target genes in a concentration-dependent manner --- p.31 / Chapter 1.4.6.1 --- Bicoid protein acts as transcriptional regulators --- p.31 / Chapter 1.4.6.2 --- Bicoid protein acts as transcriptional activators --- p.31 / Chapter 1.4.6.3 --- Bicoid protein acts as translational repressor --- p.34 / Chapter 1.5 --- Components required for the localization of bicoid mRNA --- p.35 / Chapter 1.5.1 --- Cis-acting elements --- p.35 / Chapter 1.5.1.1 --- Bicoid mRNA localization element (BLE1) at 3、UTR directs localization of bicoid mRNA --- p.36 / Chapter 1.5.2 --- Trans-acting elements --- p.37 / Chapter 1.5.2.1 --- exuperantia --- p.40 / Chapter 1.5.2.2 --- swallow --- p.41 / Chapter 1.5.2.3 --- staufen --- p.42 / Chapter 1.5.2.4 --- cytoskeleton --- p.44 / Chapter 1.6 --- Aim of project --- p.48 / Chapter CHAPTER 2 --- Characterization of exuperantia protein --- p.50 / Chapter 2.1 --- Introduction --- p.50 / Chapter 2.1.1 --- Localization step of exuperantia protein in wild type --- p.50 / Chapter 2.1.2 --- Phenotype of exuperantia mutant --- p.51 / Chapter 2.1.3 --- exuperantia gene in both female and male flies --- p.52 / Chapter 2.2 --- Materials and Methods --- p.59 / Chapter 2.2.1 --- General characteristic of exuperantia protein --- p.59 / Chapter 2.2.1.1 --- Preparation of total ovary protein from the female and male flies --- p.59 / Chapter 2.2.1.2 --- Analysis of exuperantia protein by Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis (SDS - PAGE) and Western blotting --- p.60 / Chapter 2.2.2 --- Determination of the type of phosphorylation residues in exuperantia protein --- p.61 / Chapter 2.2.2.1 --- Preparation of immunoprecipitated exuperantia protein from ovary and testis --- p.61 / Chapter 2.2.2.2 --- Dephosphorylation of exuperantia protein --- p.62 / Chapter 2.2.3 --- Two-dimensional gel electrophoresis analysis of exuperantia protein --- p.63 / Chapter 2.3 --- Results --- p.65 / Chapter 2.3.1 --- General characteristic of exuperantia protein --- p.65 / Chapter 2.3.2 --- Determination of the type of phosphorylation residues in exuperantia protein --- p.67 / Chapter 2.3.3 --- Resolving the multiple phosphorylated isoforms of exuperantia protein by two-dimensional gel electrophoresis --- p.69 / Chapter 2.4 --- Discussion --- p.72 / Chapter CHAPTER 3 --- Determination of the type of kinase(s) phosphorylate exuperantia protein --- p.77 / Chapter 3.1 --- Introduction --- p.77 / Chapter 3.2 --- Materials and Methods --- p.83 / Chapter 3.2.1 --- Phosphorylation of recombinant exuperantia protein --- p.83 / Chapter 3.2.1.1 --- Immunoprecipitation of recombinant exuperantia protein and phosphorylation reaction --- p.83 / Chapter 3.2.1.2 --- Sequential phosphorylation reaction --- p.84 / Chapter 3.2.2 --- Inhibitory effect(s) of protein kinase inhibitors on phosphorylation of native exuperantia protein --- p.85 / Chapter 3.2.2.1 --- Incubation of ovaries with protein kinase inhibitors --- p.85 / Chapter 3.2.3 --- Phosphorylation of native exuperantia protein by endogenous protein kinase(s) --- p.86 / Chapter 3.2.3.1 --- Preparation of total tissue homogenate --- p.86 / Chapter 3.2.3.2 --- Endogenous kinase assay --- p.86 / Chapter 3.3 --- Results --- p.88 / Chapter 3.3.1 --- Phosphorylation of recombinant exuperantia protein by exogenous kinase(s) --- p.88 / Chapter 3.3.2 --- Inhibitory effect(s) of protein kinase inhibitors on phosphorylation of native exuperantia protein --- p.92 / Chapter 3.3.3 --- Phosphorylation of native exuperantia protein by endogenous protein kinase(s) --- p.94 / Chapter 3.3.3.1 --- Phosphorylation of native exuperantia protein by endogenous kinase(s) with addition of protein kinase activators --- p.94 / Chapter 3.3.3.2 --- Phosphorylation of native exuperantia protein by endogenous kinase(s) with addition of protein kinase inhibitor --- p.98 / Chapter 3.4 --- Discussion --- p.101 / Chapter CHAPTER 4 --- Spatial and temporal distribution of exuperantia protein in DCO83 and exuPJ egg chambers --- p.107 / Chapter 4.1 --- Introduction --- p.107 / Chapter 4.1.1 --- Initiation of establishment of the two body axes by one single signal --- p.107 / Chapter 4.1.2 --- Stage-specific phosphorylation of exuperantia protein --- p.111 / Chapter 4.2 --- Materials and Methods --- p.113 / Chapter 4.2.1 --- Immunohistochemical distribution of exuperantia protein --- p.113 / Chapter 4.2.2 --- Stage-specific phosphorylation of exuperantia protein --- p.115 / Chapter 4.3 --- Results --- p.116 / Chapter 4.3.1 --- Immunohistochemical distribution of exuperantia protein in DCOB3 mutant --- p.119 / Chapter 4.3.2 --- Immunohistochemical distribution of exuperantia protein in exuPJ mutant --- p.121 / Chapter 4.3.3 --- Stage-specific phosphorylation of exuperantia protein in DCOB3 mutant --- p.125 / Chapter 4.3.4 --- Stage-specific phosphorylation of exuperantia protein of exuPJ mutant --- p.127 / Chapter 4.4 --- Discussion --- p.128 / Appendix A --- p.135 / Appendix B --- p.143 / References --- p.146
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_321871 |
| Date | January 1997 |
| Contributors | Cheung, Yin., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xiii, 164 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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