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姓名 吳岳隆( Yue-Long Wu)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 利用相減式雜交法鑑定小葉浮萍受巴拉刈與銅誘導之基因
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摘要(中) 本計畫是利用毒性測驗分析及分子生物技術的方法來研究及探討高等水生植物-小葉浮萍(Lemna minor )受環境化學逆境下如重金屬與有機農藥導之早期基因表達與生理反應機制。浮萍為一般常見的高等水生維管束植物,在分類上屬於Lamnaceae 此科,浮萍為單子葉植物,包含四個屬Lemna, Spirodela,Wolffiella 與Wol f f i a ,自70 年代中浮萍就常被用來作為環境污染指標與水質測驗的模式植物。而浮萍用於水質毒性測驗一般觀察葉片數目變化與葉片白化狀況。本論文比較了四種不同化學成分與作用模式的有機農藥(alachlor, m metribuzin, metsulfuron and paraquat )與重金屬(cadmium, copper, lead and zinc)對於浮萍生長與白化狀況,發現浮萍對於所處理之化學污染物皆具有敏感性,但lead 對浮萍影響不大。我們利用相減式雜交法來探討浮萍早期受paraquat 與copper 之基因表達。並進一步鑑定哪些基因受誘導。本論文在paraquat 處理的相減式cDNA 基因庫中,目前已挑出的基因,以其生理作用可分為4 群:一、解毒作用相關基因。二、抗氧化基因。三、參與二氧化碳固定作用基因。四、未知基因。在copper處理的相減式cDNA 基因庫中,目前已挑出的基因以其生理作用可分為:一、抗氧化基因。二、參與液泡運輸。三、電子傳遞。四、未知基因群。
摘要(英) The main purpose of this study is to use toxicity testing andmolecular biology approaches to study how aquatic plant duckweed,Lemna minor, responds to environmental pollutants such as heavy metalsand herbicides regarding the mechanisms of gene expression.The term “duckweed” commonly refers to a group of aquaticvascular angiosperms of the family Lamnaceae. Duckweed plants aremonocot and can be divided into four genera, Lemna, Spirodela,Wolffiella and Wolffia. Among them, Lemna minor has been frequentlyused as a model for toxicity testing with environmental pollutants sincemid 70’s. The testing can be conducted rather easily by observing theincreased frond numbers and chlorosis before and after the treatments.We compared the relative sensitivity on Lemna minor to four herbicides(alachlor, m metribuzin, metsulfuron and paraquat)and four heavy metals(cadmium, copper, lead and zinc). . Each herbicide or heavy metalrepresents distinct mode of action. The results revealed that Lemna minoris sensitive to all treatments but lead. Furthermore, we used subtraction cDNA hybridization to identify genes that are up-regulated by theselected herbicide-paraquat and heavy metals-copper.The induced genes isolated from paraquat-treated subtraction cDNA library can be divided into four groups physiologically. 1.detoxification. 2.antioxidation. 3.carbon dioxide fixation. 4.unknown. The induced genes isolated from copper treated subtraction cDNA library can be physiologically divided into three groups 1.antioxidation. 2.vesicles transport. 3. election transfer. 4. unknown .
關鍵字(中) ★ 浮萍
★ 相減式cDNA 雜交
★ 重金屬
★ 銅
★ 有機農藥
★ 巴拉刈
關鍵字(英) ★ duckweed
★ heavy metal
★ herbicide
★ paraquat
論文目次 中文摘要--------------------------------------------------------- Ⅰ
英文摘要--------------------------------------------------------- Ⅱ
目錄--------------------------------------------------------------- Ⅲ
縮寫與全名對照表--------------------------------------------- Ⅴ
圖目錄------------------------------------------------------------ Ⅵ
表目錄------------------------------------------------------------ Ⅷ
3u!B緒論-------------------------------------------------------- 1
?@!B前言---------------------------------------------------- 1
?G!B浮萍簡介---------------------------------------------- 2
?T!B毒性測驗選用之化學污染物簡介---------------- 3
¥|!B巴拉刈之特性---------------------------------------- 5
?-!B銅之特性---------------------------------------------- 6
??!B差異性基因選殖法---------------------------------- 7
?C!B研究目的---------------------------------------------- 9
L!B材料與方法------------------------------------------------ 12
?@!B浮萍來源與馴養------------------------------------- 12
?G!B重金屬與除草劑對浮萍之毒性測驗------------- 12
?T!B訊息核糖核酸 (mRNA) 之純化與分析-------- 13
¥|!B製備重金屬與有機農藥之相減式cDNA 雜交法 15
?-!B建立相減式cDNA 基因庫------------------------ 19
??!B反轉點墨法------------------------------------------- 21
?C!B基因定序與分析------------------------------------- 23
?K!B北方點墨法 ------------------------------------------ 23
?E!B化學藥品與耗材------------------------------------- 26
?Q!B儀器設備---------------------------------------------- 26.5
°N!B實驗結果-------------------------------------------------- 28
?@!B有機農藥對浮萍之毒性測驗---------------------- 28
?G!B重金屬對浮萍之毒性測驗------------------------- 28
?T!B訊息核糖核酸 (mRNA) 之純化與分析-------- 29
¥|!B有機農藥與重金屬之相減式cDNA 雜交法所
得到可能目標基因---------------------------------- 30
?-!B相減式cDNA 基因庫之誘導基因確立--------- 31
??!B反轉點墨法確立誘導基因之表現---------------- 31
?C!BDNA 定序及基因鑑定------------------------------ 33
?K!B北方墨點法鑑定單一基因------------------------- 34
﹐v!B討論-------------------------------------------------------- 36
?@!B有機農藥---------------------------------------------- 36
?G!B重金屬------------------------------------------------- 39
¥i!B建議-------------------------------------------------------- 43
3°!B參考資料-------------------------------------------------- 45
圖------------------------------------------------------------------ 52
表------------------------------------------------------------------ 90.6
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指導教授 董啟功 審核日期 2001-7-17
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