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研究生:陳凱銓
論文名稱:空腸彎曲桿菌致病因子-細胞致死腫脹毒素基因及其功能之研究分析
論文名稱(外文):Gene and functional studies of campylobacter jejuni cytolethal distending toxin
指導教授:王雯靜
學位類別:碩士
校院名稱:國立清華大學
系所名稱:生命科學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
中文關鍵詞:空腸彎曲桿菌細胞致死腫脹毒素
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彎曲桿菌為世界上最常引起人類腹瀉的致病菌,由於鑑定方法的確認及空腸彎曲桿菌基因體解讀完成,使得彎曲桿菌的研究倍受重視。目前空腸彎曲桿菌中許多致病因子中以細胞致死腫脹毒素(cytolethal distending toxin, CDT)最被研究清楚,此毒素會造成細胞腫脹、使細胞週期暫停G2/M時期、最後導致細胞死亡。在其它致病菌中也發現細胞致死腫脹毒素的存在。細胞致死腫脹毒素為一holotoxin是由三個位置相近的基因所表現,分別為cdtA、cdtB及cdtC,研究中指出三個cdt基因的表現對於細胞致死腫脹毒素的毒性是必須的,而且三個次單元會互相結合形成一tripartite complex。最近的研究發現CdtB為細胞致死腫脹毒素中負責酵素活性的次單元,CdtB的DNase I的活性會使DNA損傷,並且使細胞的細胞週期暫停。雖然對於CdtB已有較清楚的研究,但對於CdtA及CdtC的功能及在細胞致死腫脹毒素中扮演的角色仍不清楚。在本論文中,我們首先利用核酸序列來分析台灣空腸彎曲桿菌的cdt基因,並與國外的菌株比較。其次,我們利用重組His-tagged CDT蛋白質來分析CDT各個次單元的功能,由於細胞致死腫脹毒素可能為一典型的AB毒素,因此,我們以流式細胞儀來分析CDT蛋白質中的結合次單元與其結合特性。實驗結果顯示,台灣地區空腸彎曲桿菌的細胞致死腫脹毒素核酸序列與國外的核酸序列相同度達極高。另外,流式細胞儀研究結果顯CdtA次單元為CDT毒素中負責和細胞表面結合的次單元,基於上述結果,本論文推測細胞致死腫脹毒素應為一典型的AB毒素,CdtB為具有酵素活性的次單元,而CdtA或和CdtC為負責和細胞結合的次單元。

中文摘要................................................I
英文摘要.................................................II
壹、前言...........................................1
參、結果.................................................23
肆、討論..........................................................29
伍、參考文獻....................................................35
陸、圖
表...........................................................40
柒、附圖表...........................................................62

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