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論文名稱 Title |
YDL100c基因缺失對釀酒酵母在menadione下生長的影響 Effect of YDL100c Deficiency on the Growth of Saccharomyces cerevisiae in the Presence of Menadione |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
48 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2008-06-26 |
繳交日期 Date of Submission |
2008-08-01 |
關鍵字 Keywords |
釀酒酵母、氧化劑 YDL100c, menadione |
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統計 Statistics |
本論文已被瀏覽 5652 次,被下載 3 次 The thesis/dissertation has been browsed 5652 times, has been downloaded 3 times. |
中文摘要 |
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。 |
Abstract |
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. |
目次 Table of Contents |
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參考文獻 References |
1.J Silver S and Walderhaug M. Gene regulation of plasmid and chromosome-determined inorganic ion transport in bacteria. Microbiol Rev 1992, 56: 195-228. 2.Hedges, R. W. and S. Baumberg. Resistance to arsenic compounds conferred by a plasmid transmissible between strains of Escherichia coli. J.Bacteriol 1973, 115:459–460. 3.San Francisco, M. J. D., C. L. Hope, J. B. Owolabi, L. S. Tisa, and B. P.Rosen. Identification of the metalloregulatory element of the plasmidencoded arsenical resistance operon. Nucleic Acids Res 1990, 18:619–624 4.San Francisco MJ, Tisa LS, and Rosen BP. Identification of the membrane component of the anion pump encoded by the arsenical resistance operon of R-factor R733. Mol Microbiol 1989, 1:15-21. 5.Zhou T, Radaev S, Rosen B P, and Gatti D L. Structure of the ArsA ATPase:the catalytic subunit of a heavy metal resistance pump. EMBO J 2000, 19: 4838-45. 6.Mukhopadhyay R, and Rosen BP. Saccharomyces cerevisiae ACR2 gene encodes an arsenate reductase. FEMS Microbiol Lett 1998 , 168:127-36. 7.Zuniga S, Boskovic J, Jimenez A, Ballesta JP, and Remacha M. Disruption of six Saccharmyces cerevisae novel genes and phenotypic analysis of thedeletants. Yeast 1999, 15: 945-53. 8.葉瓊霞. (2001) 國立中山大學生物科學研究所碩士論文. 9.洪詩雅.(2002)國立中山大學生物科學研究所碩士論文. 10.陳涵茵.(2004)國立中山大學生物科學研究所碩士論文. 11.許展容.(2006)國立中山大學生物科學研究所碩士論文. 12.S.H. Chung, S.M. Chung, J.Y. Lee, S.R. Kim, K.S. Park, and J.H. Chung. The biological significance of non-enzymatic reaction of menadione with plasma thiols:enhancement of menadione-induced cytotoxicity to platelets by the presence of blood plasma. FEBS Lett 1999, 23: 235–240. 13.C.E. Rodriguez, M. Shinyashiki, J. Froines, R.C. Yu, J.M. Fukuto, and A.K.Cho. An examination of quinone toxicity using the yeast Saccharomyces cerevisiae model system. Toxicology 2004, 201: 185–196 14.S.H. Chung, S.M. Chung, J.Y. Lee, S.R. Kim, K.S. Park, and J.H. Chung. The biological significance of non-enzymatic reaction of menadione with plasma thiols:enhancement of menadione-induced cytotoxicity to platelets by the presence of blood plasma. FEBS Lett 1999,23: 235–240. 15.S.H. Chung, S.M. Chung, J.Y. Lee, S.R. Kim, K.S. Park, and J.H. Chung. The biological significance of non-enzymatic reaction of menadione with plasma thiols:enhancement of menadione-induced cytotoxicity to platelets by the presence of blood plasma. FEBS Lett 1999,23: 235–240. 16.J. Mauzeroll and A.J. Bard. Scanning electrochemical microscopy of menadione-glutathione conjugate export from yeast cells. Proc. Natl. Acad. Sci. U. S. A 2004,101: 7862–7867. 17.M.A. Singer and S. Lindquist. Thermotolerance in Saccharomyces cerevisiae:the Yin and Yang of trehalose. Trends Biotechnol 1998,16: 460–468. 18.Benaroudj N, Lee DH, and Goldberg AL.Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals. J Biol Chem 2001, 276:24261-7. 19.Singer MA and Lindquist S. Multiple effects of trehalose on protein foldingin vitro and in vivo. Mol Cell 1998, 1:639-48. 20.Bell W, Sun W, Hohmann S, Wera S, Reinders A, De Virgilio C, Wiemken A, and Thevelein JM.Composition and functional analysis of the Saccharomyces cerevisiae trehalose synthase complex. J Biol Chem 1998, 273:33311-9. 21.Ruis H and Schuller C. Stress signaling in yeast. Bioessays 1995, 11:959-65. 22.M.T. Martinez-Pastor, G. Marchler, C. Schuller, A.Marchler-Bauer, H.Ruis, and F.Estruch. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress-response element (STRE). EMBO J 1996, 15:2227-2235. 23.Aline de Souza Espindola, Debora Silva Gomes, Anita Dolly Panek, and Elis Cristina Araujo Eleutherio. The role of glutathione in yeast dehydration tolerance. Cryobiology 2003, 47:236-241. 24.Katrin Brombacher, Beat B. Fischer, Karin R‥ufenacht, and Rik I. L. Eggen. The role of Yap1p and Skn7p-mediated oxidative stress response in the defence of Saccharomyces cerevisiae against singlet oxygen. Yeast 2006, 23: 741–750. 25.Kuge, S., Jones, N., and Nomoto, A. Regulation of yAP-1 nuclear localization in response to oxidative stress. EMBO J 1997,16:1710-1720. 26.Vitor Costa a,b and Pedro Moradas-Ferreira a,b. Oxidative stress and Signal transduction in Saccharomyces cerevisiae:insight into ageing, apoptosis and diseases. Molecular Aspects of Medicine 2001,22:217-246. 27.Lee J, Godon C, and Lagniel G, et al. Yap1 and Skn7 control two specialized oxidative stress response regulons in yeast.J Biol Chem 1999, 274: 16040–16046. 28.Aline de Souza Espindola, Debora Silva Gomes, Anita Dolly Panek, and Elis Cristina Araujo Eleutherio. The role of glutathione in yeast dehydration tolerance. Cryobiology 2003, 47:236-241.29. 29.Jacinta A, Flattery-O'Brien and Ian W. Dawes. Hydrogen peroxide causes RAD9-dependent cell cycle arrest in G2 in Saccharomyces cerevisiae whereas menadione causes G1 arrest independent of RAD9 function . J Biol Chem 1998,15:8564-8571. 30.Singer MA and Lindquist S. Thermotolerance in Saccharomyces cerevisiae: the Yin and Yang of trehalose. Trends Biotechnol 1998,16:460-8.30. 31.Cyrne L, Martins L, Fernandes L, and Marinho HS. Regulation of Antioxidant enzymes gene expression in the yeast Saccharomyces cerevisiae during stationary phase. Free Radic Biol Med 2003, 34:385-93. 32.T.J. Chiou and W.F. Tzeng, The roles of glutathione and antioxidant enzymes in menadione-induced oxidative stress. Toxicology 2000,154: 75–84. |
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