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研究生:陳芳綺
研究生(外文):Fung-Chi Chen
論文名稱:黏菌趨化異常與發育缺陷突變株之篩尋與特性分析
論文名稱(外文):Screening and characterization of Dictyostelium mutants defective in chemotaxis and development
指導教授:陳美瑜陳美瑜引用關係
指導教授(外文):Mei-Yu Chen
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:82
中文關鍵詞:趨化運動
外文關鍵詞:chemotaxis
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趨化運動是細胞感受到化學吸引物質濃度梯度而往高濃度方向前進的過程,在很多生理與病理過程中都很重要,例如:胚胎發育、發炎、傷口癒合以及腫瘤細胞的轉移。在黏菌的整個發育過程中,趨化運動也扮演著很重要的角色。由於黏菌在實驗上操控簡單、其趨化行為與高等動物細胞十分相似,且具備多種分子研究工具,所以我們選擇以黏菌為模式系統來研究趨化運動。目前對於趨化運動的詳細機制,尚有很多不清楚的地方,為了要更加了解趨化運動的分子機轉與調控,我們希望能夠找尋跟趨化運動有關的新基因。本論文研究中,採取實驗室已經建立好的一套正向遺傳學策略,在已經建立好的REMI轉型株庫中篩尋趨化運動異常的突變株。在第一步的篩選中,我們使用兩種不同的方法,即經濾膜增集與發育型態篩檢。經濾膜增集是單純的以細胞的趨化能力來增加收集到趨化突變株的機會,而發育型態篩檢則是先找尋發育型態異常突變株之後再檢測其趨化能力並從中挑取異常者。經過這兩步驟篩尋後的突變株都必須再檢測其趨化運動的正常與否,採用的是凝膠切割及凹槽實驗兩種趨化運動測試。目前經濾膜增集流程篩檢了將近六萬個轉型株而得到22個趨化運動突變株,自發育型態篩檢流程則觀察了將近三萬個轉型株而得到8個趨化運動突變株。我對由發育型態篩檢所得的一個趨化運動突變株D15#7做了進一步分析,從中發現到一個2187 bp大小的新基因,含有一LIM domain,命名為limF。RT-PCR與北方墨點分析結果顯示此基因的表現受到發育調控,會在黏菌發育的早期表現達最高量,是否為一參與調控發育的相關基因正待研究。
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