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研究生:柯郁鈴
研究生(外文):Yu-Ling Ko
論文名稱:酵母菌Rsc58蛋白在修復DNA雙股斷裂中角色之探討
論文名稱(外文):Characterization of the Role of Yeast Rsc58p in DNA
指導教授:鄭明媛羅椀升鄭淑珍鄭淑珍引用關係
指導教授(外文):Ming-Yuan ChengWan-Sheng LuoShu-Chen Cheng
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生命科學暨基因體科學研究所
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:2009
畢業學年度:97
語文別:中文
論文頁數:57
中文關鍵詞:酵母菌雙股斷裂
外文關鍵詞:yeastRsc58double-strand break
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酵母菌Saccharomyces cerevisiae細胞中,負責染色體重塑的RSC複合物影響許多重要的生理功能,其中對於DNA雙股斷裂修復的影響,近年來已有不少證據顯示之。在RSC複合物的眾多次單位中,上有某些蛋白質的功能尚未明朗,如近年發現的Rsc58p。Rsc58p和其他RSC複合物次單位Htl1p及Swi6p存在交互作用,其胺基酸突變產物rsc58E458K是經由與Htl1p的交互作用被篩選出,且帶有rsc58E458K的突變株會有DNA致突變劑敏感的情形,因此rsc58E458K在DNA雙股斷裂修復過程中所扮演的角色值得被探討。將rsc58E458K突變合併DNA雙股斷裂修復基因刪除的菌株經由DNA致斷裂劑phleomycin感受性的測試,發現大部分菌株當rsc58E458K合併HR 修復相關基因的刪除之後,對phleomycin的感受性仍同於野生型RSC58的HR基因刪除株,雖有特定菌株被觀察到rsc58E458K/Δrad52突變株相較於Δrad52突變株有較強phleomycin抗性,且rsc58E458K/Δrad51突變株相較於Δrad51突變株有較弱phleomycin抗性,顯示對於DSB修復的影響,但經由不同菌株雜交再測試的結果發現此表型應是由特定菌株本身存在的其他未知基因突變所造成。因此,rsc58E458K對於DNA雙股斷裂的修復是否有重大影響,仍需進一步以其他方式釐清。
Saccharomyces cerevisiae RSC (Remodeling of the Structure of Chromatin) complex is known to be involved in multiple cellular processes, and is essential for yeast survival. Among the possible functions of RSC, the repair of DNA double-strand breaks (DSBs) is extremely important because improperly repaired DSBs may lead to genome instability or even cell death. Studies have shown that certain RSC subunits affect the efficiency of DSB repair. Rsc58p, a recently discovered subunit of RSC complex, has been found to interact with Htl1p and Swi6p. In a previous study, the rsc58E458K mutant also displayed sensitivity to DNA damage agents in some yeast strains. Therefore, the involvement of Rsc58p on DSB repair was investigated in this proposal. Yeast double mutants carrying rsc58E458K and the deletion of key genes involved in different DSB repair mechanisms were generated. Most strains of DSB repair mutants under the background of rsc58E458K have similar sensitivity to DNA damage agent, phleomycin, to that of wild type. However, certain rsc58E458K/Δrad52 double mutant strains suppressed the hypersensitivity to phleomycin of Δrad52 strains, and the rsc58E458K/Δrad51 double mutants were more sensitive to phleomycin than that of Δrad51 mutants. Further genetic analyses demonstrate that the phenotype is more likely caused by a non-rsc58E458K second mutation in these strains. In conclusion, the influence of rsc58E458K mutation remained obscure.
誌謝i
中文摘要ii
英iii
目錄iv
緒論1
研究目標6
材料方法7
結果15
討論20
參考文獻23
附圖27
附錄44
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