Return to search

Characterization of common carp (cyprinus carpio) insulin-like growth factor genes.

by Yu Wai Fu, Jason. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 112-120). / ACKNOWLEDGMENTS --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.iii / Chapter CHAPTER 1 --- INTORDUCTION --- p.1 / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- Historical Overview --- p.3 / Chapter 1.2.1 --- Insulin-Like Growth Factors I and II --- p.3 / Chapter 1.2.2 --- IGF Receptors --- p.5 / Chapter 1.2.3 --- IGF Binding Proteins --- p.7 / Chapter 1.3 --- Origin and Production of IGFs --- p.7 / Chapter 1.3.1 --- The Hypothalamo-Pituitary-GH-IGF-I Axis --- p.7 / Chapter 1.3.2 --- Factors Regulating IGF production --- p.9 / Chapter 1.3.3 --- Expression of IGFs in the Central Nervous System --- p.11 / Chapter 1.4 --- Actions of IGFs --- p.12 / Chapter 1.4.1 --- Insulin-like Metabolic Effects --- p.12 / Chapter 1.4.2 --- Mitogenic Effects --- p.13 / Chapter 1.4.3 --- Effects on Differentiation --- p.13 / Chapter 1.4.4 --- IGFs in Reproductive System --- p.14 / Chapter 1.4.5 --- IGF Actions in the Central Nervous System --- p.14 / Chapter 1.5 --- Transgenic and Knockout Animal Models for IGFs --- p.15 / Chapter 1.6 --- Molecular Biology of IGFs --- p.17 / Chapter 1.6.1 --- Structure of the IGF Genes --- p.17 / Chapter 1.6.2 --- Expression of IGF Genes --- p.21 / Chapter 1.6.3 --- Regulation of IGF Gene Expression --- p.23 / Chapter 1.7 --- IGF Receptors --- p.24 / Chapter 1.7.1 --- IGF-I Receptor --- p.24 / Chapter 1.7.2 --- IGF-II Receptor --- p.26 / Chapter 1.8 --- IGFBPs --- p.26 / Chapter 1.9 --- Teleost IGFs --- p.28 / Chapter 1.9.1 --- The GH-IGF-Axis in Teleost --- p.28 / Chapter 1.9.2 --- Osmoregulation and Other Biological Actions of IGFin Teleost --- p.29 / Chapter 1.9.3 --- Molecular Biology of IGFs in Teleost --- p.30 / Chapter 1.9.4 --- IGFBPs and IGF Receptors in Teleost --- p.31 / Chapter 1.10 --- Rationale and Aim of the Present Study --- p.32 / Chapter CHAPTER 2 --- SEARCH OF IGF-I PROMOTER BY GENOMIC DNA POLYMERASE CHAIN REACTION --- p.34 / Chapter 2.1 --- Introduction --- p.34 / Chapter 2.2 --- Materials --- p.35 / Chapter 2.3 --- Methods --- p.39 / Chapter 2.3.1 --- Preparation of Genomic DNA from Carp Testis --- p.39 / Chapter 2.3.2 --- Restriction Enzyme Digestion of Genomic DNA --- p.40 / Chapter 2.3.3 --- Polymerase Chain Reaction --- p.40 / Chapter 2.3.3.1 --- Ligation of the Cassette to Digested Genomic DNA --- p.40 / Chapter 2.3.3.2 --- Amplification by PCR --- p.40 / Chapter 2.3.4 --- Agarose Gel Electrophoresis --- p.42 / Chapter 2.3.5 --- Gene Clean Using Sephaglas´ёØ BandPrep Kit (Pharamica) --- p.42 / Chapter 2.3.6 --- Cloning of PCR Products --- p.43 / Chapter 2.3.7 --- Transformation of Competent Cell (Heat Shock Method) --- p.43 / Chapter 2.3.8 --- Small Scale Alkaline Preparation of Plasmid DNA --- p.44 / Chapter 2.3.9 --- Restriction Enzyme Digestion to Release the Insert --- p.45 / Chapter 2.3.10 --- Large Scale Plasmid Preparation of the Positive Clone --- p.45 / Chapter 2.3.11 --- DNA Sequencing of the Positive Clone Using the T7 DNA Polymerase Sequencing Kit (Pharmacia) --- p.46 / Chapter 2.4 --- Results and Discussion --- p.49 / Chapter CHAPTER 3 --- ISOLATION OF GENOMIC CLONES CARRYING THE IGF-I GENE --- p.55 / Chapter 3.1 --- Introduction --- p.55 / Chapter 3.2 --- Materials --- p.56 / Chapter 3.3 --- Methods --- p.58 / Chapter 3.3.1 --- Preparation of the Plating Host Cells --- p.58 / Chapter 3.3.2 --- Phage Titering --- p.58 / Chapter 3.3.3 --- Primary Screening of Common Carp Genomic Library --- p.59 / Chapter 3.3.4 --- Preparation of Radioactive Nucleic Acid Probes --- p.60 / Chapter 3.3.5 --- Purification of the Positive Clones --- p.60 / Chapter 3.3.6 --- Purification of DNA from Lambda Phage Using Sephaglas´ёØ PhagePrep Kit (Pharmacia) --- p.61 / Chapter 3.3.7 --- Restriction Enzyme Digestion Release of Inserts --- p.62 / Chapter 3.3.8 --- Capillary Transfer of DNA to Nylon Membrane Under Alkaline Condition --- p.62 / Chapter 3.3.9 --- Southern Analysis of the 10 Positive Clones --- p.63 / Chapter 3.3.10 --- Restriction Mapping of the Clone P1 --- p.64 / Chapter 3.3.11 --- Subcloning of the Fragments of the Clone PI into Plasmid Vector --- p.64 / Chapter 3.3.12 --- IGF-I Specific PCR --- p.64 / Chapter 3.3.13 --- Amplification of Introns from the Clone P1 Using PCR --- p.67 / Chapter 3.4 --- Results and Discussion --- p.70 / Chapter CHAPTER 4 --- RNA ASSAY USING REVERSE TRANSCRIPTION- POLYMERASE CHAIN REACTION --- p.83 / Chapter 4.1 --- Introduction --- p.83 / Chapter 4.2 --- Materials --- p.85 / Chapter 4.3 --- Methods --- p.86 / Chapter 4.3.1 --- Administration of Hormones --- p.86 / Chapter 4.3.1.1 --- Injection Time Course1 --- p.86 / Chapter 4.3.1.2 --- Injection Time Course2 --- p.86 / Chapter 4.3.2 --- Total RNA Extraction --- p.87 / Chapter 4.3.2.1 --- Rapid RNA Isolation --- p.87 / Chapter 4.3.3 --- Electrophoresis of RNA in Agarose Gel Containing Formaldehyde --- p.88 / Chapter 4.3.4 --- Rapid Isolation of PolyA+ mRNA from Total RNA --- p.89 / Chapter 4.3.5 --- IGF-I Specific RT-PCR --- p.90 / Chapter 4.4 --- Results and Discussion --- p.92 / Chapter CHAPTER 5 --- SEARCH FOR IGF-II GENE USING GENOMIC SOUTHERN BLOT ANALYSIS --- p.101 / Chapter 5.1 --- Introduction --- p.101 / Chapter 5.2 --- Materials --- p.103 / Chapter 5.3 --- Methods --- p.104 / Chapter 5.3.1 --- Preparation of Genomic DNA from Carp Testis --- p.104 / Chapter 5.3.2 --- Restriction Enzyme Digestion of Genomic DNA --- p.104 / Chapter 5.3.3 --- Southern Blotting of the Digested Genomic DNA --- p.104 / Chapter 5.3.4 --- Preparation of the Trout IGF-II Specific Probe --- p.104 / Chapter 5.3.5 --- Genomic Southern Hybridization --- p.105 / Chapter 5.4 --- Results and Discussion --- p.106 / Chapter CHAPTER 6 --- GENERAL DISCUSSION AND CONCLUSION --- p.109 / REFERENCES --- p.112

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_322676
Date January 1997
ContributorsYu, Wai Fu Jason., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, vii, 120 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/)

Page generated in 0.0025 seconds