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Molecular cloning and characterization of endothelin converting enzyme-2.

Ip Lai Fong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 81-92). / Abstracts in English and Chinese. / Table of Contents --- p.1 / Abbreviations --- p.4 / Chapter Chapter 1 --- Introduction and Background --- p.5 / Chapter 1.1 --- Endothelin system --- p.5 / Chapter 1.1.1 --- Endothelins --- p.5 / Chapter 1.1.2 --- Endothelin converting enzyme (ECE) isoforms --- p.12 / Chapter 1.1.3 --- Endothelin receptors --- p.24 / Chapter 1.2 --- Signal-transduction mechanisms in ET system --- p.27 / Chapter 1.3 --- The aim of the present thesis --- p.31 / Chapter Chapter 2 --- Materials and Methods --- p.32 / Chapter 2.1 --- Primer Design --- p.32 / Chapter 2.2 --- Total RNA Isolation --- p.33 / Chapter 2.3 --- Reverse transcriptase polymerase chain reaction (RT-PCR) --- p.34 / Chapter 2.3.1 --- First Strand cDNA Synthesis --- p.34 / Chapter 2.3.2 --- PCR reaction --- p.34 / Chapter 2.4 --- Agarose gel electrophoresis --- p.35 / Chapter 2.5 --- Ligation of PCR inserts to cloning vector by TA cloning method --- p.35 / Chapter 2.6 --- Competent cell preparation --- p.36 / Chapter 2.7 --- Transformation and Screening --- p.37 / Chapter 2.8 --- Plasmid DNA Extraction --- p.38 / Chapter 2.9 --- DNA sequencing --- p.38 / Chapter 2.10 --- DIG RNA Labeling --- p.38 / Chapter 2.10.1 --- Plasmid Linearization --- p.38 / Chapter 2.10.2 --- Transcription --- p.39 / Chapter 2.10.3 --- Probe purification --- p.39 / Chapter 2.11 --- In situ hybridizaion --- p.40 / Chapter 2.11.1 --- Tissue preparation and slide mounting --- p.40 / Chapter 2.11.2 --- Non-radioactive in situ hybridization --- p.41 / Chapter 2.12 --- Whole Mount non-radioactive in situ hybridization --- p.42 / Chapter 2.12.1 --- Dissection and fixation --- p.42 / Chapter 2.12.2 --- Hybridization --- p.43 / Chapter 2.12.3 --- Antibody incubation --- p.43 / Chapter 2.12.4 --- Histochemistry --- p.44 / Chapter Chapter 3 --- Results --- p.46 / Chapter 3.1 --- The molecular cloning of ECE-2 from rat brain --- p.46 / Chapter 3.2 --- Sequence characteristics of rat ECE-2 --- p.52 / Chapter 3.3 --- Comparison of rat ECE-2 with bovine and human ECE-2 and with the rat ECE-1 --- p.53 / Chapter 3.4 --- Tissue distribution of ECE-2 in rat and localization in C6 glial cells by RT-PCR --- p.60 / Chapter 3.5 --- ECE-2 in rat embryos at different gestation stages by RT-PCR --- p.60 / Chapter 3.6 --- ECE-2 distribution in C6 glioma cells --- p.63 / Chapter 3.7 --- ECE-2 distribution in rat embryo E15.5 --- p.63 / Chapter 3.8 --- ECE-2 distribution in rat brain sections --- p.63 / Chapter Chapter 4 --- p.74 / Discussion --- p.74 / References --- p.81

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323598
Date January 2001
ContributorsIp, Lai Fong., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, iv, 92 leaves : ill. ; 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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