OSTC/DC2蛋白是一個屬於人類寡醣轉移酶複合體新的次單位。寡醣轉移酶複合體在粗糙內質網的基質內，對剛形成的胜肽鏈進行N-醣化作用，鍵結上含有多甘露糖的寡醣。在此研究中，我們利用RMKLPGYLMG此序列合成的胜肽鏈當成免疫兔子的抗原，製備OSTC/DC2多株抗體。在初步的實驗結果中發現：經由熱休克後的Balb/c小鼠，在其恢復的期間，OSTC/DC2蛋白在腦部的部分組織中有增量表現的情形，且在恢復第12小時的時候，表現量達到高峰。除此之外，在細胞的熱休克模式中，OSTC/DC2的增表現也具有相同的情形。我們發現在面臨熱休克壓力下，OSTC/DC2蛋白只在大腦皮層有急遽的增量表現，顯示了OSTC/DC2可能在神經元中，有特別的功能。此外，在熱休克之後，活化的AKT及HSP70的增量表現，均與OSTC/DC2的表現相似，這顯示出OSTC/DC2在熱休克反應中，可能扮演一個重要的角色。

OSTC, a Homo sapiens OSTC/DC2 protein, belongs to a new subunit of mammalian oligosaccharyltransferase (OST). OST transfers the high-mannose sugars to nascent polypeptides during N-linked glycosylation in the lumen of the rough endoplasmic reticulum. In this study, we used synthetic peptides (RMKLPGYLMG) as an antigen to develop a polyclonal antibody against OSTC/DC2. After heat treatment of Balb/c mice, OSTC/DC2 showed up-regulated in some tissues of brain, and peaked at the 12th hour of recover stage. Furthermore, OSTC/DC2 was also up-regulated in cell lines during heat-shock treatment. The strikingly immediate up-regulation of OSTC/DC2 in response to heat shock is only seen in the cortex, suggesting that OSTC/DC2 may have unique functions in neurons. In addition, OSTC/DC2 induction is synchronous to increased expression of HSP70 and activation of AKT after heat shock, supporting the notion that OSTC/DC2 may play a significant role in heat shock response.

目 錄
論文審定書…………………………………………………………… i
誌謝…………………………………………………………………… ii
中文摘要………………………………………………………….….. iii
英文摘要………………………………………..……………………. iv

第一章 緒論………………………………………………………… 1
1-1熱休克反應…………………………………………………1
1-2蛋白質修飾…………………………………………………4
1-3醣化作用………………………………………………7
1-4酵母菌寡醣轉移酶………………………………………10
1-5醣蛋白之臨床意義………………………………………21

第二章 實驗目的……………………………………………………25

第三章 實驗材料與方法…………………………………………26
3-1生物資訊分析……………………………………………26
3-2動物與細胞………………………………………………26
3-3多株抗體製作……………………………………………29
3-4西方墨漬法……………………………………………31
3-5免疫組織化學染色………………………………………34
3-6 北方墨點法………………………………………………34
3-7 RNA干擾………………………………………………36
3-8試管內轉錄轉譯表現蛋白……………………………37
3-9熱休克處理……………………………………………37
3-10細胞轉染………………………………………38
3-11 Balb/c小鼠腦組織薄片切片…………………………39

第四章 實驗結果……………………………………………40
4-1多株抗體製備……………………………………………40
4-1-1圖表……………………………………………………44
4-2熱休克反應………………………………………………52
4-2-1圖表……………………………………………………57

第五章 討論…………………………………………………72
5-1多株抗體製備……………………………………………72
5-2熱休克反應………………………………………………74

第六章 參考文獻……………………………………………………79

第七章 附錄…………………………………………………………101

第八章 發表與得獎…………………………………………………111

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