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研究生:謝忠佑
研究生(外文):Chung-YuHsieh
論文名稱:利用原子力顯微鏡研究DNA結構變遷
論文名稱(外文):Study of DNA Structure Transition by Atomic Force Microscope
指導教授:陳宜君陳宜君引用關係
指導教授(外文):Yi-Chun Chen
學位類別:碩士
校院名稱:國立成功大學
系所名稱:物理學系碩博士班
學門:自然科學學門
學類:物理學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:52
中文關鍵詞:去氧核醣核酸原子力顯微鏡陽離子力曲線
外文關鍵詞:DNAatomic force microscopycationsforce curve
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  • 下載下載:17
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去氧核醣核酸(deoxyribonucleic acid,DNA)在生物學中是重要遺傳訊息的儲存基元。然而就目前而言,DNA還有許多未知特性需要被研究探討。本實驗利用雲母和金作為基板,以原子力顯微鏡(atomic force microscopy, AFM)研究吸附於基板的DNA結構。
雲母和DNA都帶有負電荷,可以藉由多價陽離子作為橋樑使DNA固定於雲母基板。於實驗中藉由改變離子濃度改善DNA吸附狀況。當以乙醇取代去離子水作為溶劑時,觀察DNA結構轉變。金和DNA鹼基都具有疏水性會產生強力鍵結,可以利用探針去拉伸同時鍵結金和探針的DNA,量測DNA的力曲線(force curve)。
DNA吸附在雲母上的效果不完全會隨陽離子濃度增加而提升,陽離子濃度介於0.5 mM ~ 10 mM時效果最好。當DNA溶於乙醇結構會由B構形轉變為A構形。量測DNA力曲線時探針的速度會影響所量測的曲線,當受力達到27 pN時DNA的螺旋狀結構會因為拉伸而解開。
Deoxyribonucleic acid (DNA) is the most important element for storing genetic information in biology. Until so far, there are still a lot of unknown properties which need to be investigated. In this study, we used mica and gold substrate for binding DNA and observed DNA structures by using atomic force microscopy (AFM).
Both mica and DNA have negative charges, so multivalent cations can be the bridge to link DNA and mica substrate. In our experiments, cation concentration is changed as the parameter to improve the DNA binding efficiency. We also observed the transition of DNA conformation when the liquor of deionized water was replaced by ethanol. Both Gold and the base on DNA backbone are hydrophobic, so DNA can be bound to gold substrate strongly. As the result, we used the tip to stretch DNA that is both bound to the gold substrate and the tip, and then we can measure the force curve of DNA during structure transformation.
The efficiency of DNA bound on mica is not really proportional to cation concentration. There are the best efficiency when the cation concentration is between 0.5 mM ~ 10mM. The conformation of DNA will change from B-form to A-form . During the stretching of DNA, The velocity of tip will affect the experiment data of force curve. The helical conformation of DNA will be unwinding under stretching with 27 pN.
摘要...................................................... I
Abstract..................................................II
誌謝.....................................................III
總目錄.....................................................IV
表目錄......................................................VI
圖目錄.................................................... VII
第一章 緒論................................................ 1
第二章文獻回顧 ............................................. 2
2.1 去氧核醣核酸............................................ 2
2.1.1 核苷酸結構與性質....................................... 2
2.1.2 DNA結構............................................. 4
2.2 基板特性............................................... 9
2.2.1 雲母................................................ 9
2.2.2 金和DNA的交互作用.....................................11
第三章 實驗原理與方法....................................... 13
3.1 掃描式探針顯微鏡........................................ 13
3.1.1 掃描式探針顯微鏡的原理與架構............................ 13
3.1.2 ScanAsyst掃描模式................................... 18
3.2 實驗流程.............................................. 21
3.2.1 固定Lambda-DNA於雲母基板............................. 21
3.2.2 固定DNA(Poly(dA))於金基板.............................22
第四章 實驗結果與討論....................................... 24
4.1 DNA與雲母基板的作用情形................................. 24
4.1.1 鎳離子濃度對於DNA吸附的影響............................ 27
4.1.2 醇類對DNA結構的影響................................... 33
4.1.3 DNA物理性質的量測.................................... 35
4.2 Poly(dA)力曲線(Force curve)的量測...................... 43
第五章結論................................................ 49
參考資料.................................................. 50
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