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  1. NTU Theses and Dissertations Repository
  2. 生命科學院
  3. 漁業科學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30759
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor周宏農
dc.contributor.authorJian-Zhi Linen
dc.contributor.author林健智zh_TW
dc.date.accessioned2021-06-13T02:14:45Z-
dc.date.available2007-05-24
dc.date.copyright2007-05-24
dc.date.issued2007
dc.date.submitted2007-04-27
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30759-
dc.description.abstract本研究論文分為兩大主題,一為探討利用定量PCR技術,檢測水體中微囊藻族群數的可行性;另一實驗則觀察本實驗室一株由桃園單離培養的微囊藻株,其藻毒合成與釋放的相互關係。
實驗一,藉由設計七組不同產毒基因片段(mcyA ~ E and J及mcy promoter)之引子,並以台灣地區單離培養的微囊藻種(8株產毒,四株不產毒)作為定量PCR最佳檢測目標基因的測試。定量PCR結果顯示:(1)所有產毒微囊藻皆具有mcys,而不產毒微囊藻則含有微量或不含有mcys;(2)PCR產物進行鎔點分析,發現mcyD的序列最為保守,適合作為產毒微囊藻的定量目標基因;(3)微囊藻細胞數與定量PCR的Ct值呈直線相關;(4)定量PCR的分析結果不受野外樣品與實驗室藻混合樣品的影響。定量PCR具備有快速且靈敏的優點,相當適合用於環境微囊藻族群的監測之用。
實驗二,過去的研究相信,微囊藻毒主要存在於細胞中,只有當細胞老化或死亡時,才會將毒素釋出,然本實驗室發現一特殊的微囊藻株M.TY-1,此藻株在指數生長期時具有大量藻毒釋放的現象。藉由McyH蛋白質序列比對,發現其與不釋放藻毒的藻株僅一個胺基酸發生突變,由於McyH被認為有藻毒釋放與合成藻毒的雙重功能,因此M.TY-1將可提供為後續藻毒釋放研究的模式品系。
zh_TW
dc.description.abstractThe suitability of using mcy genes analysis for screening toxic Microcystis populations and the release of microcystins were determined. Seven characteristic segments, mcyA~E, J, and the promoter of the microcystin synthetase gene cluster, designed in a Q-PCR amplification were applied for the quantitative measurement of toxic populations in environmental samples. Observations during the method-development experiments against 8 toxic and 4 non-toxic clones of Microcystis were: (i) the expected specific amplicons were found in all toxic clones but absent or less amount in non-toxic clones; (ii) all the toxic clones showed consistent Tm values of mcyD, implying this partial mcyD segment to be the most conserved one among tested mcys; (iii) a linear correlation was obtained between the microscopically determined cell numbers and the PCR threshold cycles; (iv) cell concentration of the toxic Microcystis from Q-PCR measurement was not affected by field sample and the addendum of non-toxic populations. Q-PCR is sensitivity and provides an efficient technology suitable for toxic Microcystis population screening.
Microcystins are considered primary intracellular and microcystins release was generally considered to be linked to a decrease in the integrity of the cells. However, from a strain of M. aeruginosa designated as M.TY-1, it was found that microcystins are released during the log phase of growth and continuously accumulated in the cell-free medium. A single amino acid difference of the sequence McyH was found in the toxin-releasing strain in comparison with other non-toxin-releasing strains. Sequence of McyH was known to be homologous to the bacteria ABC transporter, but also known as a key protein that maintained the integrity of microcystin synthetase clusters. This significant toxin release strain would be a model for the research of the M. aeruginosa toxin release system.
en
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Previous issue date: 2007
en
dc.description.tableofcontents摘要---------------------------------------------------------------------------ii
壹、 文獻回顧------------------------------------------------------------- 1
貳、 實驗一、開發以定量PCR 進行產毒微囊藻族群計算可行性探討
一、 目的------------------------------------------------------------- 30
二、 前言------------------------------------------------------------- 30
三、 材料方法------------------------------------------------------- 34
四、 結果------------------------------------------------------------- 40
五、 討論------------------------------------------------------------- 50
參、 實驗二、微囊藻株M.TY-1 藻毒釋放觀察
一、 目的------------------------------------------------------------- 55
二、 前言------------------------------------------------------------- 55
三、 材料方法------------------------------------------------------- 57
四、 結果------------------------------------------------------------- 59
五、 討論------------------------------------------------------------- 66
肆、 參考文獻------------------------------------------------------------ 69
伍、 附錄------------------------------------------------------------------ 77
陸、 個人發表------------------------------------------------------------ 91
dc.language.isozh-TW
dc.title微囊藻產毒基因檢測技術開發與微囊藻毒素釋放生理研究zh_TW
dc.titleConcerns of the application of mcy genes analysis in the toxic Microcystis population screening and observations on the release of microcystinsen
dc.typeThesis
dc.date.schoolyear95-2
dc.description.degree碩士
dc.contributor.oralexamcommittee宋賢一,黃穰,黃鵬鵬
dc.subject.keyword微囊藻,微囊藻毒,定量PCR,藻毒釋放,zh_TW
dc.subject.keywordMicrocystis,Microcystins,Real-Time PCR,Toxin release,en
dc.relation.page92
dc.rights.note有償授權
dc.date.accepted2007-04-27
dc.contributor.author-college生命科學院zh_TW
dc.contributor.author-dept漁業科學研究所zh_TW
顯示於系所單位:漁業科學研究所

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