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研究生:李佩怡
研究生(外文):Pei-I Li
論文名稱:二價錳離子對耐輻射奇異球菌蛋白體表現的影響
論文名稱(外文):Effects of manganese on the proteomic expression in Deinococcus radiodurans
指導教授:劉仲康劉仲康引用關係
指導教授(外文):Jong-Kang Liu
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:2002
畢業學年度:90
語文別:中文
論文頁數:215
中文關鍵詞:耐輻射奇異球菌
外文關鍵詞:Deinococcus radiodurans
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二價錳離子(Mn2+)會誘導耐輻射奇異球菌(Deinococcus radiodurans) 細胞再次分裂,產生二次生長,此現象稱為Mn-CD (Mn-induced Cell Division) 效應,而此加錳離子誘導後之新生細胞,體積較母細胞小,其抗輻射能力較母細胞差、且紅色色素生成量也少很多;而其他一些與抗氧化有關的酵素,例如超氧離子歧化酉每(superoxide dismutase)及觸酉每(catalase)的活性則有增加的情形。而其他二價金屬離子則無法造成Mn-CD效應。本研究是利用二維蛋白質電泳(2-D Electrophoresis)的方式與Matrix Assisted Laser Desorption Ionization (MALDI) MASS分析法,來分析本菌在有或無錳離子培養下,其菌體內基因表現出的蛋白質種類與數量的差異。結果發現在添加錳離子培養的菌體有明顯被抑制的蛋白質為ribosomal protein L20、L35、L34、chain 3 of the large ribosomal subunit、preprotein translocase (SecE subunit)以及三種hypothetical proteins (DR1423、DR1897、一基因編號未知)。ribosomal protein與菌體內蛋白質的合成有直接相關性,顯示錳離子可能影響細胞蛋白質的合成。此外,加入錳離子培養的菌體亦會誘發acetyl-CoA acetyltransferase、transcriptional regulator (TetR family)、phosphinothricin acetyltransferase,以及三種hypothetical proteins (DR0214、DR0296、DRA0100)的表現,但這些hypothetical proteins的功能還未知。這些現象證明了錳離子確實可誘導本株細菌產生不同程度的蛋白體表現,以及利用這些表現出來的蛋白質來進行不同的生理代謝途徑。


The addition of Mn2+ 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 this study, 2-D electrophoresis and Matrix Assisted Laser Desorption Ionization (MALDI) MASS were used to analyze the proteomic differences between Mn-CD and normal uninduced cells. The results showed that the expression of ribosomal proteins L20, L34, L35, chain 3 of the large ribosomal subunit, preprotein translocase (SecE subunit) and three hypothetical proteins (DR1423, DR1897 and one unidentified gene product), were pressed significantly in the cell culture when Mn2+ were added. Since these ribosomal proteins are responsible for the synthesis of protein, it is clear that Mn2+ could disturb the proteins synthesis in this bacterium. The addition of Mn2+could also induce the expression of acetyl-CoA acetyltransferase, transcriptional regulator (TetR family), phosphinothricin acetyltransferase and three hypothetical proteins (DR0214, DR0296 and DRA0100). The functions of these hypothetical proteins were not known yet. In conclusion, it is true that Mn2+ could alter the proteomic expression and some metabolic pathways in D. radiodurans.


中文摘要………………………………………………..…………I
英文摘要…………………………………………………………..II
表目錄……………………………………………………………..III
圖目錄……………………………………………………………...V
壹、前言………………………………………………………..1
貳、材料方法…………………………………………………..25
參、結果…………………………………………………………33
肆、討論…………………………………………………………39
伍、參考文獻……………………………………………………45
陸、圖表…………………………………………………………58


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