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研究生:黃淑君
研究生(外文):Shu-Jiun Huang
論文名稱:YDL100c基因缺失對釀酒酵母在menadione下生長的影響
論文名稱(外文):Effect of YDL100c Deficiency on the Growth of Saccharomyces cerevisiae in the Presence of Menadione
指導教授:許清玫
指導教授(外文):Ching-Mei Hsu
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物科學系研究所
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:2008
畢業學年度:96
語文別:中文
論文頁數:48
中文關鍵詞:釀酒酵母氧化劑
外文關鍵詞:YDL100cmenadione
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本實驗研究釀酒酵母YDL100c基因缺失對酵母在添加50 μM menadione後生長的影響,初步結果顯示,添加menadione時YDL100c基因缺失的KO菌株較WT菌株的生長落後,且KO菌株細胞內分子的氧化程度及細胞膜脂質的過氧化程度均較WT菌株增加,此與KO菌株的catalase活性在添加menadione後較WT菌株活性低一致,顯示YDL100c基因缺失時,menadione會對細胞造成更嚴重的傷害。進一步測定細胞內非酵素性抗氧化成分GSH與trehalose的含量時,結果顯示添加menadione後KO菌株的GSH含量比WT菌株減少,顯示YDL100c基因缺失時,菌株細胞在氧化壓力下可能無法誘發GSH1的表現以進行GSH的合成,而添加menadione後KO菌株的trehalose含量較WT菌株增加,顯示trehalose含量與KO菌株生長落後有關。而在抗氧化相關基因如CTT1、SOD1、GSH1、TPS1、TRR1與TRX2的表現方面,在添加menadione後,除了屬於general stress response的TPS1和CTT1基因的表現減少外,屬於specific stress response的GSH1、SOD1、TRR1及TRX2也下降。綜合以上結果推測,釀酒酵母在menadione存在時,因應氧化壓力而調控general stress response與specific stress response的表現,但YDL100c基因的缺失可能影響兩者的啟動機制而造成KO菌株的生長落後甚至死亡。
Wild type strain (WT) and YDL100c disrupted strain (KO) were grown at 30oC for 8 hr after adding 50 μM menadione. Cells of both strains were assayed for trehalose accumulation, intracellular molecular oxidation level, membrane lipid oxidation, and glutathione (GSH) content. The data showed both the molecular and membrane lipid oxidation levels are higher and the GSH content is lower in KO compared with WT in the presence of menadione. The catalase activity in KO strain are reduced than that in WT strain, catalase activity would affect in the presence of menadione. Further study of antioxidant gene expressions showed that TPS1 and CTT1 were involved in the general stress response;SOD1, GSH1, TRR1 and TRX2 were involved in the specific stress response. Above mentioned mRNA level were reduced, suggesting that the deletion of YDL100c in S. cerevisiae affects the operation of general and specific stress response when grown in the presence of menadione.
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材料與方法----------------------------------------------------------------11
結果-------------------------------------------------------------------------18
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表----------------------------------------------------------------------------30
討論-------------------------------------------------------------------------31
參考文獻-------------------------------------------------------------------36
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附表-------------------------------------------------------------------------44
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