Ho, Sin Chu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 112-135). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.ii / Acknowledgement --- p.iii / Table of Contents --- p.iv / List of Abbreviations --- p.vii / List of Figures --- p.x / List of Tables --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Test Organisms: Daphnia magna and Moina macrocopa --- p.1 / Chapter 1.2 --- Biomarkers --- p.2 / Chapter 1.3 --- Heat shock protein (HSP) --- p.4 / Chapter 1.3.1 --- Regulation of hsp70 gene expression --- p.5 / Chapter 1.3.2 --- Review of studies on hsp70 of various species --- p.6 / Chapter 1.3.3 --- Review of studies using molecular methods in Daphnia and Moina --- p.7 / Chapter 1.4 --- Environmental stress studied --- p.9 / Chapter 1.5 --- Objectives --- p.11 / Chapter Chapter 2 --- Materials and Methods --- p.12 / Chapter 2.1 --- Experimental organisms --- p.12 / Chapter 2.2 --- "The effects of temperature, metals and nutritional stress on cladocerans" --- p.13 / Chapter 2.3 --- Demographic measurement --- p.15 / Chapter 2.4 --- Cloning and sequencing of D. magna and M. macrocopa hsp70 --- p.17 / Chapter 2.4.1 --- Acute heat shock --- p.17 / Chapter 2.4.2 --- Total RNA extraction --- p.17 / Chapter 2.4.3 --- Reverse transcription and polymerase chain reaction (PCR) --- p.18 / Chapter 2.4.4 --- Degenerative PCR --- p.21 / Chapter 2.4.5 --- Cloning and sequencing of hsp70 partial sequence --- p.25 / Chapter 2.4.6 --- Rapid amplification of cDNA ends (RACE) --- p.28 / Chapter 2.4.7 --- Confirmation of D. magna and M. macrocopa hsp70 sequences --- p.30 / Chapter 2.5 --- Molecular measurements - heat shock protein 70 gene expression --- p.31 / Chapter 2.5.1 --- Sample collection --- p.31 / Chapter 2.5.2 --- Quantification of cDNA level by real-time PCR --- p.32 / Chapter 2.5.2.1 --- Primer design --- p.32 / Chapter 2.5.2.2 --- Validation of real-time PCR conditions --- p.32 / Chapter 2.5.2.3 --- Determination of hsp70 gene expression levels --- p.35 / Chapter 2.6 --- Statistical analysis --- p.36 / Chapter Chapter 3 --- Results --- p.37 / Chapter 3.1 --- Demographic measurements --- p.37 / Chapter 3.1.1 --- Effects of temperature on life history parameters of M macrocopa --- p.37 / Chapter 3.1.2 --- Effects of temperature on life history parameters of D. magna --- p.40 / Chapter 3.1.3 --- Effects of Metals on life history parameters of M. macrocopa --- p.42 / Chapter 3.1.4 --- Effects of Anabaena on life history parameters of M. macrocopa --- p.47 / Chapter 3.2 --- Cloning and sequencing of hsp70 --- p.50 / Chapter 3.2.1 --- Partial sequences of D. magna and M. macrocopa hsp70 --- p.50 / Chapter 3.2.2 --- Sequences of 3´ة and 5´ة RACE amplified products --- p.53 / Chapter 3.2.3 --- Full sequences of D. magna and M. macrocopa hsp70 --- p.53 / Chapter 3.2.4 --- Phylogenetic relationships of the hsp70 gene --- p.60 / Chapter 3.3 --- Molecular measurements - heat shock protein 70 gene expression --- p.71 / Chapter 3.3.1 --- Validation of real-time PCR conditions --- p.71 / Chapter 3.3.2 --- Effect of temperature stress on hsp70 gene expression levels --- p.76 / Chapter 3.3.3 --- Effect of metals on hsp70 gene expression levels --- p.78 / Chapter 3.3.4 --- Effect of Anabaena on Hsp70 Gene Expression Levels --- p.83 / Chapter Chapter 4 --- Discussion --- p.85 / Chapter 4.1 --- Demographic responses of M. macrocopa and D. magna by different stress --- p.85 / Chapter 4.1.1 --- Effects of temperature on life history parameters --- p.85 / Chapter 4.1.2 --- Effects of metal stress on life history parameters --- p.87 / Chapter 4.1.3 --- Effects of Anabaena on life history parameters --- p.89 / Chapter 4.2 --- Characterization of hsp70 of M. macrocopa and D. magna --- p.92 / Chapter 4.3 --- Hsp70 gene expression in response to different stress --- p.94 / Chapter 4.3.1 --- Effect of temperature stress on hsp70 gene expression levels --- p.94 / Chapter 4.3.2 --- Effect of metals on Hsp70 gene expression levels --- p.97 / Chapter 4.3.3 --- Effect of Anabaena on hsp70 gene expression levels --- p.101 / Chapter 4.4 --- Comparison of demographic and molecular measurements --- p.102 / Chapter 4.5 --- Performance of hsp70 gene expression as a biomarker of environmental stress --- p.106 / Chapter 4.6 --- Conclusion --- p.111 / References --- p.112 / Appendix --- p.136