by Ying Tat Mak. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 178-198). / Title --- p.1 / Abstract --- p.2 / Acknowledgments --- p.5 / Contents --- p.6 / Abbreviations --- p.9 / List of Tables --- p.11 / List of Figures --- p.13 / Chapter Chapter 1: --- Introduction / Chapter 1.1 --- Cholesterol Metabolism and Atherosclerosis --- p.15 / Chapter 1.1.1 --- Cholesterol and Cholesterol Metabolism --- p.17 / Chapter 1.1.2 --- Cholesterol Transport: Apolipoprotein and Lipoprotein --- p.23 / Chapter 1.1.3 --- Cholesterol and Atherosclerosis --- p.26 / Chapter 1.2 --- Hyperlipidaemia --- p.30 / Chapter 1.2.1 --- Primary and Secondary Hyperlipidaemia --- p.31 / Chapter 1.2.2 --- Mutations leading to Primary Hypercholesterolaemia --- p.36 / Chapter 1.3 --- Familial Hypercholesterolaemia --- p.38 / Chapter 1.3.1 --- Historical Aspects --- p.38 / Chapter 1.3.2 --- Clinical Features - Diagnosis and Consequences --- p.39 / Chapter 1.3.3 --- Population Prevalence --- p.40 / Chapter 1.3.4 --- Mutations in the Low Density Lipoprotein Receptor Gene --- p.41 / Chapter 1.4 --- Methods for Detecting Mutations in LDL Receptor Gene --- p.51 / Chapter 1.4.1 --- Southern Blotting Based Methods --- p.51 / Chapter 1.4.2 --- Polymerase Chain Reaction Based Methods --- p.52 / Chapter 1.4.3 --- Screening Methods for Unknown Mutations in LDL Receptor Gene --- p.56 / Chapter 1.5 --- Mutations of the LDL receptor gene in Chinese --- p.58 / Chapter Chapter 2: --- Objectives --- p.63 / Chapter Chapter 3: --- Materials and Methods / Chapter 3.1 --- Subjects / Chapter 3.1.1 --- Familial Hypercholesterolaemia Patients --- p.65 / Chapter 3.1.2 --- Normocholesterolaemia Subjects --- p.67 / Chapter 3.2 --- Materials / Chapter 3.2.1 --- Enzymes --- p.67 / Chapter 3.2.2 --- DNA Markers --- p.68 / Chapter 3.2.3 --- Reagents Kits --- p.68 / Chapter 3.2.4 --- Primers for PCR --- p.68 / Chapter 3.2.5 --- Chemicals and Reagents --- p.69 / Chapter 3.2.6 --- Radioisotopes --- p.70 / Chapter 3.2.7 --- Solutions and Buffers --- p.70 / Chapter 3.3 --- Methods / Chapter 3.3.1 --- Blood Collection --- p.71 / Chapter 3.3.2 --- General Biochemistry Tests --- p.72 / Chapter 3.3.3 --- DNA Extraction --- p.72 / Chapter 3.3.4 --- RNA Extraction --- p.73 / Chapter 3.3.5 --- Polymerase Chain Reaction --- p.74 / Chapter 3.3.6 --- Agarose Gel Electrophoresis --- p.76 / Chapter 3.3.7 --- Polyacrylamide Gel Electrophoresis --- p.78 / Chapter 3.3.8 --- Single Strand Conformation Polymorphism --- p.79 / Chapter 3.3.9 --- Reverse Transcription - Polymerase Chain Reaction --- p.79 / Chapter 3.3.10 --- Direct DNA Sequencing --- p.81 / Chapter 3.3.11 --- Haplotyping of the LDL receptor gene --- p.83 / Chapter 3.3.12 --- Restriction Enzyme Digestion --- p.84 / Chapter Chapter 4: --- Results / Chapter 4.1 --- Patients Investigations --- p.88 / Chapter 4.1.1 --- Normal Control Subjects --- p.88 / Chapter 4.1.2 --- Patients --- p.88 / Chapter 4.2 --- PCR-SSCP Analysis of LDL Receptor Gene --- p.90 / Chapter 4.3 --- Summary of Mutations Identified --- p.92 / Chapter 4.4 --- Novel Mutations --- p.94 / Chapter 4.5 --- Previously Reported Mutations --- p.97 / Chapter 4.6 --- Polymorphisms and Silent Mutation --- p.100 / Chapter 4.6.1 --- New Polymorphism --- p.100 / Chapter 4.6.2 --- New Silent Mutation --- p.102 / Chapter 4.6.3 --- Reported Polymorphisms --- p.103 / Chapter 4.7 --- Southern Blotting --- p.103 / Chapter 4.8 --- Haplotypes --- p.104 / (All Figures for Chapter 4) --- p.106 / Chapter Chapter 5: --- Discussions / Chapter 5.1 --- Use of SSCP in Screening for Mutations and Polymorphisms --- p.158 / Chapter 5.2 --- Novel and Reported Mutations --- p.160 / Chapter 5.3 --- Novel Polymorphism and Silent Mutation --- p.170 / Chapter 5.4 --- Common Polymorphisms --- p.171 / Chapter 5.5 --- Possible Common Mutations of the LDL Receptor Gene in Chinese --- p.172 / Chapter 5.6 --- Pattern of LDL Receptor Gene Mutations in Chinese --- p.173 / References --- p.178