利用蛋白質體學分析四逆湯對耐輻射奇異球菌生長之影響

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Title:	利用蛋白質體學分析四逆湯對耐輻射奇異球菌生長之影響 Proteomic analysis of a Si Ni Tang-induced Growth in Radioresistant Bacterium Deinococcus radiodurans
Authors:	翁嘉禪 Chia-shan Weng
Contributors:	鄭淨月嘉南藥理科技大學：生物科技研究所
Keywords:	耐輻射奇異球四逆湯二維蛋白質電泳基質輔助雷射脫附游離質譜儀 Matrix Assisted Laser Desorption Ionization Mass 2-D electrophoresis Si ni tang Deinococcus radiodurans Mn-induced Cell Division (Mn-CD)
Date:	2008
Issue Date:	2008-12-29 15:21:33 (UTC+8)
Abstract:	Mn-induced Cell Division (Mn-CD) 效應會誘導耐輻射奇異球菌(Deinococcus radiodurans) 二次生長。誘導後之新生細胞被發現與抗氧化有關的酵素，例如超氧離子歧化酶(superoxide dismutase)及觸酶(catalase)的活性則有增加的情形；此效應是其他二價金屬離子所無法造成的。因此在本研究中，添加四逆湯於培養液中觀察其是否能促進耐輻射奇異球菌之生長，及比較蛋白質表現量和未添加前有無差異性。實驗方法是利用二維蛋白質電泳(2-D Electrophoresis) 和基質輔助雷射脫附游離質譜儀(Matrix Assisted Laser Desorption Ionization Mass Spectrometry；MALDI/MS)來分析耐輻射奇異球菌在有或無錳離子與四逆湯的培養下，其菌體表現出的蛋白質種類與數量的差異。實驗結果發現當添加錳離子與四逆湯進行培養時，有多種蛋白質被誘導出來，表現量增加；有兩種蛋白質被抑制，其中已鑑定出phosphopyruvate hydratase是為被誘導表現的蛋白質，為參與糖解作用中的催化酵素之一，顯示錳離子與四逆湯可能影響菌體的糖解作用。此外也鑑定出另二種被誘導表現的conserved hypothetical protein，但其弁鄐揖憚鴃C另外，實驗也發現某一hypothetical protein(其弁鄐揖憚?在添加錳離子時被抑制，在加入四逆湯時表現增加。根據以上結果證明四逆湯的添加除了能和錳離子產生相同效果外，亦能產生不同程度的蛋白質表現。 The addition of Mn (Ⅱ) to stationary phase culture of Deinococcus radiodurans could induce further cell division. This type of cell division termed Mn-induced Cell Division (Mn-CD).The Mn-CD cells were less resistant to radiation, having smaller cell size, and forming less red-pigment. However, the activities of antioxidation enzymes such as superoxide dismutase (SOD) and catalase, were increased. No other divalent metal ions could cause Mn-CD. In addition, the addition of Si ni tang to culture of D.radiodurans could induce cell growth and protein expression. In this study, 2-D electrophoresis and Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI/MS) were used to analyze the proteomic differences between adjunction of Mn (Ⅱ) and Si ni tang in normal cells. The results showed that the expression of phosphopyruvate hydratase was induced when Mn (Ⅱ) and Si ni tang were added. The phosphopyruvate hydratase is one of catalase in glycolysis. It is showed Mn (Ⅱ) and Si ni tang to probably influence the process of glycolysis. The addition of Mn (Ⅱ) and Si ni tang could induce two conserved hypothetical protein were increased. The functions of conserved hypothetical protein were not known yet. The expression of hypothetical protein was induced when Si ni tang added and deceased when Mn (II) added. The function of hypothetical protein was not known yet. In conclusion, the results showed that Si ni tang produced a growth effect as same as the Mn II and induced the difference in the expression of protein.
Relation:	校內校外均不公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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