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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 廖秀娟 | |
| dc.contributor.author | Yueh-Fen Li | en |
| dc.contributor.author | 李悅芬 | zh_TW |
| dc.date.accessioned | 2021-06-08T05:11:47Z | - |
| dc.date.copyright | 2006-07-29 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-21 | |
| dc.identifier.citation | Abernathy CO, Liu YP, Longfellow D, Aposhian HV, Beck B, Fowler B, Goyer R, Menzer R, Rossman T, Thompson C, and Waalkes M. 1999. Arsenic: hear effects, mechanisms of actions, and research issues. Environ Health Perspect 107: 593-7.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23857 | - |
| dc.description.abstract | The environmental pollution threatens human health as well as wildlife in the past decades. As a result, it is necessary to develop sensitive, effective, and inexpensive methods which can efficiently monitor and determine the presence and amount of hazards in the environment. In this study, we report two biologically analytical methods to detect and measure the environmental pollutants.
Firstly, we describe the construction and characterization of the fluorescent and the luminescent Escherichia coli whole-cell biosensors for the detection of bioavailable toluene and its related compounds. The biosensor strains Escherichia coli DH5α carrying pTOLGFP or pTOLLUX were developed based on the expression of reporter genes: gfp or luxCDABE under the control of the Pu promoter and xylR gene of Pseudomonas putita plasmid pWW0. To assess their applicability for analyzing environmentally relevant samples, the biosensor harboring pTOLLUX was field-tested on water and soil samples collected from toluene contaminated sites. Our results demonstrate that nonpathogenic bacterial biosensors developed in the present study is useful and applicable in determining the bioavailability of toluene and its related compounds with high sensitivity in environmental samples, and they suggest a potential for its inexpensive application in field-ready tests. Secondly, alterations of glutathione levels as well as the mRNA levels of HSP70 in tilapia fish were investigated under arsenite exposure. Tilapia fish were exposed to waterborne arsenite (0, 1, 2, and 4 ppm, respectively) for 1 day and 3 day exposure duration. After the treatment, arsenite concentrations in gill, intestine, liver, and muscle were measured by means of inductively coupled plasma (ICP). Meanwhile, the GSH contents in fish tissues were measured and the mRNA expression of HSP70 was analyzed with semi-quantitative RT-PCR. Our results indicate that HSP70 mRNA expression and GSH levels exhibited a correlation with the arsenite exposure condition as well as the arsenic accumulation, indicating their usefulness as the biomarker of arsenite exposure in tilapia. Furthermore, the bioaccumulations of arsenic differentially distributed in various fish tissues assist in providing information of bioavailability. We conclude that biological assays, such as bacterial biosensors and biomarker presented in this study, for detecting and determining the environmental contaminants can compensate the disadvantages of traditional analytical methods such as not capable to reflect the bioavailability and failing to show the effects of contaminants mixtures on organisms, and thus providing critical data that can be useful in risk assessment. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T05:11:47Z (GMT). No. of bitstreams: 1 ntu-95-R93622013-1.pdf: 2752575 bytes, checksum: 2bdac91f99b99a338dbff18612b5a196 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 致 謝 I
中文摘要 III ABSTRACT V TABLE OF CONTENTS VII LIST OF TABLES IX LIST OF FIGURES X ABBREVIATIONS XI CHAPTER 1 INTRODUCTION 1 Overview of environmental pollution 1 1.1 Bacterial biosensor of toluene and its related compounds 2 1.1.1 Toluene 2 1.1.2 Regulation of xyl operon 3 1.1.3 Bacterial biosensor 6 1.2 Biomarker of arsenic exposure 9 1.2.1 Arsenic 9 1.2.2 Biomarker 11 1.2.3 Tilapia 12 1.2.4 HSP70 12 1.2.5 Glutathione 13 1.3 Purposes of study 15 1.3.1 Bacterial biosensor of toluene and its related compounds 15 1.3.2 Biomarker of arsenite exposure 16 CHAPTER 2 MATERIALS AND METHODS 17 Chemicals 17 2.1 Bacterial biosensor of toluene 17 2.1.1 Construction of biosensor plasmids 17 2.1.2 Bacteria Cultivation and induction experiments 19 2.1.3 Measurements of GFP fluorescence and luminescence in culture 21 2.1.4 Microscopic detection of fluorescent biosensor 22 2.1.5 Selectivity studies of biosensors 22 2.1.6 Kinetic analysis of GFP fluorescence and luminescence induced by toluene 23 2.1.7 Testing of contaminated soil and water samples with bacteria biosensors 23 2.1.8 Data analysis 24 2.2 Biomarker of arsenic exposure 24 2.2.1 Test organisms 24 2.2.2 Treatment of fish 25 2.2.3 Isolation of total RNA from fish 25 2.2.4 Reverse transcription and polymerase chain reaction (RT-PCR) 26 2.2.5 Chemical analysis of arsenic 28 2.2.6 Intracellular GSH measurement 28 CHAPTER 3 RESULTS 30 3.1 Bacterial biosensor of toluene and its related compounds 30 3.1.1 Construction of the bacterial biosensors 30 3.1.2 Selectivity analysis of biosensor to toluene-related compounds 30 3.1.3 Time-dependent induction of biosensor with effector 33 3.1.4 Dose-dependent induction of biosensor with effector 35 3.1.5 Testing of contaminated water and soil with the biosensor 37 3.2 Biomarker of arsenic exposure 41 3.2.1 Arsenic accumulation in tilapia 41 3.2.2 HSP70 as biomarker of arsenic exposure 41 3.3.3 GSH as biomarker of arsenic exposure 43 CHAPTER 4 DISCUSSION 50 4.1 Bacterial biosensor of toluene and its related compounds 50 4.1.1 Development of bacterial biosensors of toluene 50 4.1.2 Characterization of bacterial biosensors of toluene 50 4.1.3 Comparison of the bacterial biosensor assay with the traditional analysis method 52 4.1.4 Comparison of reporter protein in biosensor 54 4.1.5 Field testing of environmental samples using a biosensor approach 56 4.2 Biomarker of arsenic exposure 57 4.2.1 Arsenic accumulation in tilapia 57 4.2.2 HSP70 mRNA expression as biomarker of arsenic exposure 58 4.2.3 GSH as biomarker of arsenic exposure 59 CHAPTER 5 CONCLUSION 61 5.1 Bacterial biosensor for toluene and its related compounds 61 5.2 Biomarker of the arsenic exposure 61 REFERENCE 62 APPENDIX 73 | |
| dc.language.iso | en | |
| dc.title | 利用生物方法檢測環境中的污染物 | zh_TW |
| dc.title | Biological assays for the detection and measurement of environmental pollutants | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 沈偉強,李心予 | |
| dc.subject.keyword | 細菌生物偵測器,甲苯,生物獲取率,砷,生物指標, | zh_TW |
| dc.subject.keyword | Bacterial biosensors,Toluene,Bioavailability,Arsenic,Biomarker, | en |
| dc.relation.page | 73 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2006-07-22 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物環境系統工程學研究所 | zh_TW |
| 顯示於系所單位： | 生物環境系統工程學系 | |
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