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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 溫進德(Jin-Der Wen) | |
dc.contributor.author | Tsung-Huan Wu | en |
dc.contributor.author | 吳宗桓 | zh_TW |
dc.date.accessioned | 2021-06-17T08:19:44Z | - |
dc.date.available | 2024-08-22 | |
dc.date.copyright | 2019-08-22 | |
dc.date.issued | 2019 | |
dc.date.submitted | 2019-08-13 | |
dc.identifier.citation | Ashkin, A. (1970). 'Acceleration and Trapping of Particles by Radiation Pressure.' Physical Review Letters 24(4): 156-159.
Budworth, H. and C. T. McMurray (2013). 'A brief history of triplet repeat diseases.' Methods in molecular biology (Clifton, N.J.) 1010: 3-17. Carlos Bustamante, Yann R. Chemla, a. Nancy R. Forde and D. Izhaky (2004). 'Mechanical Processes in Biochemistry.' Annual Review of Biochemistry 73(1): 705-748. Jain, A. and R. D. Vale (2017). 'RNA phase transitions in repeat expansion disorders.' Nature 546: 243. Levinson, G. and G. A. Gutman (1987). 'Slipped-strand mispairing: a major mechanism for DNA sequence evolution.' Molecular Biology and Evolution 4(3): 203-221. Liphardt, J., B. Onoa, S. B. Smith, I. Tinoco and C. Bustamante (2001). 'Reversible Unfolding of Single RNA Molecules by Mechanical Force.' Science 292(5517): 733-737. Manosas, M., J. D. Wen, P. T. X. Li, S. B. Smith, C. Bustamante, I. Tinoco, Jr. and F. Ritort (2007). 'Force unfolding kinetics of RNA using optical tweezers. II. Modeling experiments.' Biophysical journal 92(9): 3010-3021. Nalavade, R., N. Griesche, D. P. Ryan, S. Hildebrand and S. Krauß (2013). 'Mechanisms of RNA-induced toxicity in CAG repeat disorders.' Cell Death &Amp; Disease 4: e752. Paulson, H. (2018). 'Repeat expansion diseases.' Handbook of clinical neurology 147: 105-123. Richards, R. I. (2001). 'Dynamic mutations: a decade of unstable expanded repeats in human genetic disease.' Human Molecular Genetics 10(20): 2187-2194. Samadashwily, G. M., G. Raca and S. M. Mirkin (1997). 'Trinucleotide repeats affect DNA replication in vivo.' Nature Genetics 17(3): 298-304. Smith, S. B., Y. Cui and C. Bustamante (1996). 'Overstretching B-DNA: The Elastic Response of Individual Double-Stranded and Single-Stranded DNA Molecules.' Science 271(5250): 795-799. Smith, S. B., Y. Cui and C. Bustamante (2003). [7] Optical-trap force transducer that operates by direct measurement of light momentum. Methods in Enzymology, Academic Press. 361: 134-162. Zlatanova, J. and K. van Holde (2006). 'Single-Molecule Biology: What Is It and How Does It Work?' Molecular Cell 24(3): 317-329. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/74095 | - |
dc.description.abstract | 重複序列的擴增會影響DNA複製、轉錄、RNA結構以及蛋白質異常結合、聚集。然而序列和結構的不穩定性,使我們難以透過傳統巨量的生物化學方式來進行研究。在本研究中,透過單分子技術,我們想了解CAG重複序列的摺疊結構與展開方式,並與普通的雙股螺旋比較。利用光鉗,將帶有雙股DNA handles的CAG重複序列連結在兩個珠子之間,以機械應力展開其結構再放鬆使其摺疊。結果顯示,在力-延展量的作圖中,CAG重複序列呈現明顯的「高原區」,力的大小相對延展量增加並無太大改變。顯示CAG重複序列在打開或摺疊中的每一小步中不斷的局部重組,同時CAG重複序列最終能夠形成極為緊密的結構。此外,CAG重複摺疊與打開呈現了準可逆過程的特徵,與雙股螺旋結構解開的不可逆過程形成強烈對比。但是較高的離子濃度也會升高CAG重複序列的結構強度,降低可逆性。從固定光阱的實驗也佐證了CAG重複序列的結構具有結構的多型性。本研究揭示了CAG重複序列的摺疊結構與潛在的分子致病機制,可能協助未來能針對其特異的結構特性找到應用,或是其疾病的治療方式。 | zh_TW |
dc.description.abstract | Trinucleotide expansion can affect DNA replication, transcription, RNA structure and abnormal protein binding or aggregation. Due to the instability, the structure is difficult to study by traditional bulk biochemical experiments. In this study, we apply the single molecule techniques to study the structure of CAG repeats and how they fold and unfold. On optical tweezers, DNA handles with CAG repeats are tethered to beads for mechanical unfolding. Here, we show the force-extension plots of the CAG repeats exhibit a distinct force plateau during unfolding. Our results suggested that CAG repeats form stable, compact structures which unfold and refold in continual substeps. Moreover, such unfolding/refolding resembled as a quasi-reversible process, contrary to irreversible unfolding of Watson-Crick helices. We also found high concentrations of metal ions also increase the structural strength of the CAG repeats and reduce the reversibility. This study reveals the structure and the potential pathogenic mechanism of CAG repeats. The characteristic feature of the repeats may be used in medical treatment or other applications. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T08:19:44Z (GMT). No. of bitstreams: 1 ntu-108-R06b43029-1.pdf: 2482729 bytes, checksum: db9b467947a86c444a01df9c6f050791 (MD5) Previous issue date: 2019 | en |
dc.description.tableofcontents | 中文摘要 i
ABSTRACT ii 圖目錄 v 第一章 導論 1 1.1 去氧核醣核酸 1 1.2 三核苷酸重複序列擴增 1 1.3 結構多型性與擴增序列不穩定性 2 1.4 單分子技術 2 1.5 光鉗 Optical tweezers 2 1.6 研究動機 3 第二章 材料與方法 4 2.1 材料 4 2.1.1 質體 4 2.1.2 引子 4 2.1.3 試劑及藥品 4 2.1.4 酵素 5 2.1.5 實驗溶液 5 2.1.6 實驗用樣品 6 2.2 方法 6 2.2.1 聚合酶連鎖反應 6 2.2.2 光鉗實驗 7 2.2.3 機械性展開CAG重複序列結構 8 2.2.4 固定光阱的動態觀察 8 2.2.5 資料分析 9 第三章 結果 10 3.1 用PCR製作QXX樣品 10 3.2 機械性展開CAG重複序列結構 10 3.2.1 Q100P 10 3.2.2 Q20P 11 3.2.3 Q60P 12 3.3 鎂離子對CAG重複序列結構的影響 12 3.3.1 Q100P 13 3.3.2 Q20P 13 3.3.3 Q60P 13 3.4 慢速度展開 14 3.4.1 Q100P 14 3.4.2 Q20P 14 3.4.3 Q60P 15 3.5 固定光阱CAG重複序列動態觀察 15 3.5.1 Q100P 16 3.5.2 Q20P 17 第四章 討論 18 4.1 CAG重複序列對複製的影響 18 4.2 CAG重複序列的結構 18 4.3 鎂離子穩定結構降低可逆性 19 4.4 慢速展開與固定光阱實驗 20 4.5 定力分子(constant-force molecular) 21 4.6 未來展望 21 參考文獻 23 | |
dc.language.iso | zh-TW | |
dc.title | CAG三核苷酸重複序列擴增的單分子力學研究 | zh_TW |
dc.title | Single Molecule Mechanical Study of Expanded CAG Trinucleotide DNA Repeats | en |
dc.type | Thesis | |
dc.date.schoolyear | 107-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張功耀(Kung-Yao Chang),周信宏(Hsin-Hung David Chou) | |
dc.subject.keyword | 光鉗,重複序列擴增,結構多型性,序列不穩定性,聚麩醯胺酸疾病, | zh_TW |
dc.subject.keyword | optical tweezers,expanded repeats,structural polymorphism,repeat instability,polyglutamine diseases, | en |
dc.relation.page | 52 | |
dc.identifier.doi | 10.6342/NTU201903311 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2019-08-14 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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