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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 孔繁璐 | |
| dc.contributor.author | Ting-Yu Chen | en |
| dc.contributor.author | 陳亭妤 | zh_TW |
| dc.date.accessioned | 2021-06-13T03:35:17Z | - |
| dc.date.available | 2011-08-03 | |
| dc.date.copyright | 2006-08-03 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-07-27 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32177 | - |
| dc.description.abstract | 阿茲海默氏症屬於一種神經退化疾病，有兩種重要特徵:其一是神經纖維糾結，另一則是老年斑。老年斑的主要成分為Aβ，是amyloid precursor protein (APP) 經過一連串酵素序列性切割作用後的產物。因此，了解APP的代謝過程對於預防及治療阿茲海默氏症相當重要。
我們之前以酵母雙雜交實驗找出了一些可與APP的C端 (AICD) 產生交互作用的人類腦蛋白質，其中之一是一個在腦部組織有大量表現的lipid raft marker，flotillin-1。以一株神經母細胞瘤細胞株 (SH-SY5Y) 的蛋白質萃取液進行免疫共沉澱實驗，證實了APP C端片段與flotillin-1之間確實有associated，並非direct interaction之證據。此外，我們利用一些flotillin-1的deletion mutants，試圖藉由酵母雙交實驗和免疫共沉澱實驗，找出flotillin-1上與AICD作用、以及與其他flotillins間形成聚合體相關的重要區域。實驗結果顯示，一個位於flotillin-1 N端的putative oligomerization domain (137-188 a.a.) 對其與AICD之交互作用是重要的。另一個位於flotillin-1 C端的putative oligomerization domain (189-282 a.a.) 則是對其產生聚合體是重要的。另外，有關APP的磷酸化與flotillin-1形成聚合體之間關係的實驗正在進行中。 | zh_TW |
| dc.description.abstract | Alzheimer’s disease (AD) is a neurodegenerative disease. Two hallmarks of AD are neurofibrillary tangles (NFTs) and senile plaques (SPs). The main component of SPs is Aβ, the product of a serial enzymatic digestion of amyloid precursor protein (APP). Therefore, it is important to understand the processing of APP to prevent or cure AD.
Human brain proteins interacting with APP-intracellular-C-terminal-domain (AICD) were identified using a yeast two-hybrid screening assay. One of the protein candidates that may interact with AICD is flotillin-1, a lipid raft marker expressed predominantly in brain tissues. The association between APP and its C-terminal-fragment (CTF) with flotillin-1 was confirmed by co-immunoprecipitation experiment using the protein extracts from a neuroblastoma cell line SH-SY5Y. In addition, a series of deletion mutants of flotillin-1 were generated to identify region(s) on flotillin-1 important for the interaction between AICD and flotillin-1 or between flotillins by yeast two-hybrid assay and co-immunoprecipitation experiment. Our result suggests that a putative oligomerization domain near the N-terminus of flotillin-1 (137-188 a.a.) may be crucial for the AICD-flotillin-1 interaction. The other putative oligomerization domain near the C-terminus of flotillin-1 (189-282 a.a.) may be crucial for flotillin-1-flotillin-1 interaction. Experiments to elucidate whether phosphorylation of APP is associated with oligomerization of flotillin-1 are still ongoing. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T03:35:17Z (GMT). No. of bitstreams: 1 ntu-95-R93423004-1.pdf: 1501306 bytes, checksum: e6228f3af4ca4ea5f869742709546db7 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 中文摘要 1
Abstract 2 英文縮寫表 3 序論 6 實驗目的 15 材料與方法 16 A. 確認APP CTF和flotillin-1之間的交互作用 16 1. 培養SH-SY5Y細胞 16 2. 抽取蛋白質與蛋白質定量 17 3. 免疫共沉澱 17 3.1 製備50% protein A beads slurry 17 3.2 連接protein A beads與抗體 17 3.3 Cross-link protein A beads與抗體 18 3.4抗體與蛋白質萃取液作用 18 4. SDS-PAGE樣本製備 18 5. SDS-PAGE (tris-tricine gel) 19 6. 西方墨點法 19 B. 找出並確認flotillin-1上可與AICD作用的區域、找出並確認flotillin-1上可與其形成聚合體的區域 20 1. 製備質體 20 2. 洋菜膠電泳與純化限制酶切割產物 20 3. Ligation 和transformation 20 4. 篩選 21 5. Co-transformation與酵母雙雜交 21 6. 確認質體 22 7. 抽取蛋白質與蛋白質定量 23 8. 免疫共沉澱 23 9. SDS-PAGE樣本製備 23 10. SDS-PAGE (tris-glycine gel) 23 11. 西方墨點法 24 C. 探討APP磷酸化與flotillin-1形成聚合體之間的關聯 24 1. 培養SH-SY5Y細胞 24 2. APP的誘發磷酸化與抑制磷酸化 24 3. SDS-PAGE樣本製備 25 4. SDS-PAGE (tris-tricine、tris-glycine gels) 25 5. 西方墨點法 25 結果與討論 26 結論 30 圖與表 31 參考文獻 45 | |
| dc.language.iso | zh-TW | |
| dc.title | APP C端片段與flotillin-1間交互作用之研究 | zh_TW |
| dc.title | Investigation of the Interaction between APP-C-terminal-fragment and Flotillin-1 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 顧記華,兵岳忻,李財坤,林榮信 | |
| dc.subject.keyword | 阿茲海默氏症, | zh_TW |
| dc.subject.keyword | flotillin-1,APP,lipid raft, | en |
| dc.relation.page | 52 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-07-27 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 藥學研究所 | zh_TW |
| 顯示於系所單位： | 藥學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-95-1.pdf 目前未授權公開取用 | 1.47 MB | Adobe PDF |
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