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Thesis (Ph.D.)--University of Melbourne, Dept. of Anatomy and Cell Biology, 2008. / Typescript. Includes bibliographical references (leaves 235-267)

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Siu-wai Michael Sung. / Thesis submitted in: December 2000. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 97-111). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abbreviations --- p.v / Table of Contents --- p.vii / Chapter Chapter 1 --- General Introduction / Chapter l. 1 --- Drosophila as a model for studying development --- p.1 / Chapter l .2 --- The formation of the body axis in Drosophila --- p.2 / Chapter l .3 --- Maternal genes are essential for development --- p.9 / Chapter 1.4 --- Maternal gene bicoid is essential for formation of the anterior structures in the embryo --- p.13 / Chapter 1.5 --- Establishment of an anterior to posterior bicoid protein gradient --- p.13 / Chapter 1.6 --- The bicoid protein gradient controls the downstream zygotic target genes in a concentration-dependent manner --- p.17 / Chapter 1.7 --- Bicoid protein acts as transcriptional regulators \9 --- p.19 / Chapter 1.8 --- Bicoid protein acts as transcriptional regulators --- p.21 / Chapter 1.9 --- The anterior localization of bcd mRNA --- p.21 / Chapter 1.10 --- Components required for bcd mRNA localization at anterior pole of oocyte / Chapter 1.10.1 --- Cis-acting elements --- p.22 / Chapter 1.10.1.1 --- BLE1 at 3' UTR directs localization of bcd mRNA --- p.23 / Chapter 1.10.2 --- Trans-acting elements / Chapter 1.10.2.1 --- "Exuperantia, swallow, and staufen are necessary for localization for bcd mKNA" --- p.27 / Chapter 1.10.2.2 --- exu protein is an absolute requirement for localization for bcd mRNA --- p.30 / Chapter 1.10.2.3 --- Microtubules dependence of localization --- p.31 / Chapter 1.11 --- Functions of exu in localization of bcd mRNA --- p.32 / Chapter 1.12 --- Characteristics of Bicoid protein and Bic-D gene --- p.33 / Chapter 1.13 --- Aim of Project --- p.36 / Chapter CHAPTER 2 --- Materials and Methods / Chapter 2.1 --- Fly Food --- p.37 / Chapter 2.2 --- Conditions in maintaining the fly stocks and working stocks --- p.37 / Chapter 2.3 --- Localization of exu protein and other intracellular elements by indirect immunofluorescence detection / Chapter 2.3.1 --- Immunohistrochemical distribution of exu and Bic-D protein --- p.38 / Chapter 2.3.2 --- Immunohistrochemical distribution of β-tubulin --- p.39 / Chapter 2.4 --- Preparation of total protein from the female and male flies --- p.41 / Chapter 2.5 --- Analysis of interactions between exu and trans-acting elements / Chapter 2.5.1 --- 35S-methionine metabolic labelling and immunoprecipitation by RIPA buffer --- p.41 / Chapter 2.5.2 --- 35S-methionine metabolic labelling and immunoprecipitation by Mach and Lehmann buffer system --- p.43 / Chapter 2.6 --- Co-immunoprecipitation of exu and Bic-D protein / Chapter 2.6.1 --- Co-immunoprecipitation of exu and Bic-D protein synthesized by in vitro coupled transcription and translation system with modified Mach and Lechmann buffer system --- p.44 / Chapter 2.7 --- in vivo ovary extract co-immunoprecipitation / Chapter 2.7.1 --- in vivo ovary extraction co-immunoprecipitation of exu and Bic-D protein with modified Mach and Lehmann buffer system supplemented with recombinant exu protein --- p.45 / Chapter CHAPTER 3 --- Results / Chapter 3.1 --- Analysis of co-localization of exu and Bic-D protein by double immuno-fluorescence staining on w1118 flies --- p.47 / Chapter 3.2 --- Analysis of co-localization of exu protein and β-tubulin protein by double immuno-fluorescence staining on w1118 flies --- p.51 / Chapter 3.3 --- Analysis of co-localization of exu and Bic-D protein by double immuno-fluorescence staining on Bic-D mutants --- p.55 / Chapter 3.4 --- Co-immunoprecipitation of exu and Bic-D protein synthesized by in vitro coupled transcription and translation system --- p.61 / Chapter 3.5 --- 35S-Methionine metabolic labelling and co-immunoprecipitation of exu and Bic-D protein with RIP A buffer system --- p.65 / Chapter 3.6 --- 35S-Methionine metabolic labelling and co-immunoprecipitation of exu and Bic-D protein with Mach and Lehmann buffer system --- p.68 / Chapter 3.7 --- in vivo ovary extract co-immunoprecipitation of exu and Bic-D protein with modified Mach and Lehmann buffer system supplemented with recombinant exu protein --- p.71 / Chapter CHAPTER 4 --- Discussion / Chapter 4.1 --- Analysis of co-localization of exu protein and other intracellular elements by indirect double immunofluorescence staining detection --- p.74 / Chapter 4.2 --- Analysis of co-localization of exu and BicD protein by double immuno- fluorescence staining on Bic-D mutants --- p.78 / Chapter 4.3 --- Co-immunoprecipitation of exu and BicD protein synthesized by in vitro coupled transcription and translation system --- p.79 / Chapter 4.4 --- Analysis of interactions between exu and trans-acting elements by 35S- Methionine metabolic labelling and immunoprecipitation --- p.82 / Chapter 4.5 --- "in vivo ovary extract coimmunoprecipitation of exu and Bic-D protein with modified Mech and Lehmann buffer system, supplemented with recombinant exu protein" --- p.84 / Chapter 4.6 --- Recent developments on the concept of ribonucleoprotein --- p.86 / Appendix A Supplementary protocols --- p.91 / Appendix B Reagents --- p.95 / Reference --- p.97