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Heavy metal contamination and metallothionein mRNA levels in the tissues of tilapia.

Lam Kwok Lim. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 107-126). / Abstract also in Chinese. / Acknowledgments --- p.i / Presentations Derived from the Present Thesis Work --- p.ii / Abstract --- p.iv / Abbreviations --- p.vii / Abbreviation Table for Amino Acids --- p.ix / List of Figures --- p.x / List of Tables --- p.xii / Contents --- p.xiii / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Metallothionein (MT) --- p.1 / Chapter 1.1.1 --- Classification of MT --- p.1 / Chapter 1.1.2 --- Structure of MT --- p.2 / Chapter 1.1.3. --- Structure of MT Genes --- p.4 / Chapter 1.1.4 --- Function of MT --- p.5 / Chapter 1.1.5 --- Regulation of MT Expression --- p.7 / Chapter 1.1.6 --- Fish MT --- p.9 / Chapter 1.1.7. --- Aims and Rationale of the Present Study --- p.12 / Chapter 2 --- MT mRNA Induction of Tilapia After Intraperitoneal Injection of Metal --- p.18 / Chapter 2.1 --- Introduction --- p.18 / Chapter 2.1.1. --- Specific Aims of This Chapter --- p.19 / Chapter 2.2 --- Materials and Methods --- p.20 / Chapter 2.2.1 --- Regents --- p.20 / Chapter 2.2.1.1 --- Purification of Total RNA --- p.20 / Chapter 2.2.1.2 --- Denaturing Gel and Vacuum Blotting of RNA (Northern Blotting) --- p.20 / Chapter 2.2.1.3 --- Hybridization --- p.21 / Chapter 2.2.2 --- Methods --- p.21 / Chapter 2.2.2.1 --- Purification of Total RNA --- p.21 / Chapter 2.2.2.2 --- Vacuum Blotting of Total RNA (Northern Blotting) --- p.22 / Chapter 2.2.2.3 --- Radioactive Labeling of Nucleic Acid Probes --- p.22 / Chapter 2.2.2.4 --- Hybridization --- p.22 / Chapter 2.2.2.5 --- Densitometric Analysis --- p.23 / Chapter 2.2.2.6 --- Calculation of MT mRNA Levels and Analysis of Results --- p.23 / Chapter 2.2.3 --- Endogenous MT mRNA Expression of Juvenile Tilapia and Carp --- p.23 / Chapter 2.2.4 --- Induction of MT mRNA Juvenile Tilapia and Carp Injected with Metals --- p.24 / Chapter 2.3 --- Results --- p.25 / Chapter 2.3.1 --- Endogenous Levels of MT mRNA in Tilapias in Normal Conditions --- p.25 / Chapter 2.3.2 --- Induction of MT mRNA Levels in Juvenile Tilapia Injected with Metals --- p.25 / Chapter 2.3.1.1 --- Copper Injection --- p.25 / Chapter 2.3.1.2 --- Zinc Injection --- p.25 / Chapter 2.3.1.3 --- Cadmium Injection --- p.26 / Chapter 2.3.3 --- Induction of MT mRNA Levels in Juvenile Carp with Zinc Injection --- p.26 / Chapter 2.4 --- Discussion --- p.26 / Chapter 2.4.1 --- MT mRNA Expression of Tilapia and Carp Injected with Metals --- p.26 / Chapter 2.5 --- Conclusions --- p.29 / Chapter 3 --- Induction Level of MT mRNA in Tilapia After Aqueous Exposure to Metals --- p.35 / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.1.1 --- Specific aims of this chapter --- p.36 / Chapter 3.2 --- Material s and Methods --- p.36 / Chapter 3.2.1 --- 96hours LC-50 values for zinc and copper --- p.36 / Chapter 3.2.2 --- Induction of MT mRNA in Juvenile Tiapias under Metal Aqueous Exposures --- p.37 / Chapter 3.2.3 --- Calculation of Fold Induction of MT mRNA and Analysis of Results --- p.38 / Chapter 3.2.4 --- Metal Analysis --- p.38 / Chapter 3.3 --- Results --- p.38 / Chapter 3.3.1 --- LC-50 values of metals for Juvenile Tilapia --- p.38 / Chapter 3.3.2 --- Induction of MT mRNA in Juvenile Tilapia under Metal Aqueous Exposures --- p.39 / Chapter 3.3.2.1 --- Aqueous Exposure to Copper --- p.39 / Chapter 3.3.2.2 --- Aqueous Exposure to Zinc --- p.40 / Chapter 3.3.2.3 --- Aqueous Exposure to Cadmium --- p.41 / Chapter 3.3.3 --- Induction of MT mRNA in Juvenile Carp after Aqueous Exposures to Metal --- p.41 / Chapter 3.3.3.1 --- Aqueous Exposure to Cadmium --- p.41 / Chapter 3.3.4 --- Metal Concentrations of Water Samples from the Aquaria in the Metal Exposure Test of Tilapia and Carp --- p.42 / Chapter 3.4 --- Discussion --- p.42 / Chapter 3.4.1 --- LC-50 values of Metals for Tilapia --- p.42 / Chapter 3.4.2 --- MT mRNA Expression of Tilapias under Metal Aqueous Exposure --- p.44 / Chapter 3.4.3 --- Normalization of the Signals of Northern Blot Analysis --- p.47 / Chapter 3.5 --- Conclusions --- p.48 / Chapter 4 --- Field Study --- p.58 / Chapter 4.1 --- Introduction --- p.58 / Chapter 4.1.1 --- Specific Aims of this Chapter --- p.59 / Chapter 4.2 --- Materials and Methods --- p.59 / Chapter 4.2.1 --- Sampling Sites --- p.59 / Chapter 4.2.2 --- Data Analysis --- p.60 / Chapter 4.2.3 --- Harvest of Feral Tilapia --- p.60 / Chapter 4.2.4 --- Determination of Metal Concentration of Metal Concentration in the Tissues of Feral Tilapia --- p.60 / Chapter 4.2.5 --- Endogenous MT mRNA Levels Using Northern Blot Analysis --- p.61 / Chapter 4.2.6 --- Calculation of MT mRNA Levels and Analysis of Results --- p.61 / Chapter 4.3 --- Results --- p.62 / Chapter 4.3.1 --- Metal Concentrations in the Tissues of Feral Tilapia --- p.62 / Chapter 4.3.2 --- Comparison of Metal Concentrations Among Different Tissues of Feral Tilapia --- p.62 / Chapter 4.3.3 --- MT mRNA Levels in the Tissues of Feral Tilapia --- p.63 / Chapter 4.3.4 --- Correlation Between Metal Concentrations and Endogenous MT mRNA Levels in the Tissues of Feral Tilapia --- p.63 / Chapter 4.4 --- Discussion --- p.64 / Chapter 4.4.1 --- Bioaccumulation of Metals --- p.64 / Chapter 4.4.2 --- Endogenous Levels of MT mRNA in the Feral Tilapia --- p.67 / Chapter 4.5 --- Conclusions --- p.68 / Chapter 5 --- Cloning of Tilapia MT Genes --- p.86 / Chapter 5.1 --- Specific Aims of This Chapter 、 --- p.86 / Chapter 5.2 --- Materials and Methods --- p.87 / Chapter 5.2.1 --- Regents --- p.87 / Chapter 5.2.1.1 --- Preparation of Plasmid DNA --- p.87 / Chapter 5.2.1.2 --- Preparation of Genomic DNA --- p.87 / Chapter 5.2.1.3 --- Restriction Enzyme Digestion --- p.88 / Chapter 5.2.1.4 --- Vacuum Blotting of DNA (Southern Blotting) --- p.88 / Chapter 5.2.1.5 --- Polymerase Chain Reaction --- p.89 / Chapter 5.2.1.6 --- Transformation of E.coli Competent Cells --- p.89 / Chapter 5.2.1.7 --- Nucleotide Sequence Determination --- p.89 / Chapter 5.2.1.8 --- List of Primers --- p.90 / Chapter 5.2.1.8.1 --- Primers for Nucleotide Sequence Determination --- p.90 / Chapter 5.2.1.8.2 --- Tilapia MT Specific Primers for PCR --- p.90 / Chapter 5.2.2 --- Methods --- p.91 / Chapter 5.2.2.1 --- Preparation of Plasmid --- p.91 / Chapter 5.2.2.2 --- Preparation of Genomic DNA --- p.91 / Chapter 5.2.2.3 --- Preparation of Enzyme Digestion --- p.92 / Chapter 5.2.2.4 --- Vacuum Blotting of Genomic DNA (Southern Blotting) --- p.92 / Chapter 5.2.2.5 --- Radioactive Labeling of Nucleic Acid Probes --- p.92 / Chapter 5.2.2.6 --- Hybridization --- p.93 / Chapter 5.2.2.7 --- Polymerase Chain Reaction --- p.93 / Chapter 5.2.3 --- Southern Blot Analysis of Tilapia Genomic DNA --- p.93 / Chapter 5.2.4 --- Analysis of the Sequences of Tilapia MT Genes --- p.94 / Chapter 5.2.4.1 --- Amplification of MT Genes Using PCR --- p.94 / Chapter 5.2.4.2 --- Cloning of the MT Genes --- p.94 / Chapter 5.2.4.3 --- Transformation of E.coli Competent Cell --- p.94 / Chapter 5.2.4.4 --- Nucleotide Sequence Determination --- p.95 / Chapter 5.3 --- Results --- p.95 / Chapter 5.3.1 --- Southern Blot Analysis of Tilapia Genomic DNA --- p.95 / Chapter 5.3.2 --- Amplification of MT Gene Fragments Using PCR --- p.95 / Chapter 5.3.3 --- Analysis of the Sequences of Tilapia MT Genes --- p.96 / Chapter 5.4 --- Discussion --- p.96 / Chapter 5.4.1 --- Fish MT Genes --- p.96 / Chapter 5.5 --- Conclusions --- p.98 / Chapter 6 --- General Discussion --- p.104 / References --- p.107

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_322357
Date January 1998
ContributorsLam, Kwok Lim., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvii, 126 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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