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研究生:劉恬均
研究生(外文):Tian-Jiun Liu
論文名稱:系統性分析細菌突變株的表面增強拉曼散射光譜以呈現出菌體套膜上特殊結構之拉曼光譜特徵
論文名稱(外文):Systemic analyses of SERS profiles in bacterial mutants reveal Raman spectral features that represent specific envelope components
指導教授:林奇宏林奇宏引用關係
指導教授(外文):Chi-Hung Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生醫光電工程研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:90
中文關鍵詞:拉曼
外文關鍵詞:raman
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  • 下載下載:74
  • 收藏至我的研究室書目清單書目收藏:0
細菌的表層結構,和其他生物體一樣,是其與外界環境相互連繫的管道,它們在許多生理功能上皆扮演著重要角色:如保護菌體不受外界物質傷害、感知外界刺激、吸收養份、感染宿主及運動等,所以研究此表層結構將有助於我們了解更多有關菌體對於外界反應的相關知識並解決其致病性的問題。

利用表面增強拉曼散射平台所得到的拉曼光譜可以提供我們更多有關生物體表面分子結構組成的資訊 ,是一非常適合用以研究微生物細胞壁表面組成成份的工具。但目前連結光譜資料與生物體結構組成兩者之間尚缺乏完整的資料庫與技術平台。

在此研究中,我們利用大腸桿菌K-12的一系列套膜內組成的單一基因剔除突變株,來嘗試連結光譜資料與菌體表層特異結構組成,系統性分析光譜資料得到的樹狀結構圖結果,提供了一個以結構相似度分類的依據,並以基因轉植的方式將突變株缺失的基因送回,得到與野生株相似的拉曼光譜,證明了套膜上特殊結構在拉曼光譜上的特徵。
The structure of bacterial surface serves as the bridge between cell and environment. It plays an important role in many physiological functions such as protecting bacteria from surrounding materials, sensing stimulations, intake nutrition, infection and motility. Studying the components of bacterial surface helps us to understand more about their responses to environment and solve the pathogenesis problems.

The Raman spectra provided by the Surface enhanced Raman scattering (SERS) platform provides us much information about the molecular composition of the cell surface structure. It is very suitable tool in researching the composition of microbial cell wall surface structure. However, currently it is a lack of the complete database and technique platform for linking the spectrum data and the cell surface components.

To connect the spectral information and bacterial surface specific component, we used a set of the E. coli K12 single gene knockout mutants. Based on spectral similarity, the dendrogram derived from the hierarchical clustering provides a preliminary way for identifying the single gene knockout mutants.
目錄
圖表目錄
Abstract
中文摘要
第一章 緒論
1.1 研究動機細菌表層結構
1.2 細菌表層結構
1.3 套膜內主要成份構造
1.4 格蘭氏陰性細菌套膜內特殊結構
第二章 理論基礎
2.1 拉曼散射歷史背景
2.2 拉曼效應(Raman effect)
2.3 共振拉曼效應
2.4 表面增強拉曼散射效應
2.5 表面增強拉曼散射在生物醫學領域上之應用
第三章、材料與方法
3.1 細菌菌株 (Bacterial strain)
3.2 引子與質體 (Plasmid and primers)
3.3 抗體 (Antibodies)
3.4 細菌的培養 (Bacterial culture)
3.5 細菌之質體萃取與純化 (Plasmid extraction)
3.6 瓊脂凝膠製作及電泳 (Agarose gel electrophoresis)
3.7 聚合酶連鎖反應 (polymerase chain reaction; PCR)
3.8 DNA 之萃取與純化 (DNA purification)
3.9 重組DNA的選殖 (Cloning)
3.10 純化帶有6xHis tag的重組蛋白質 (Protein Purification)
3.11 SDS-凝膠電泳 (SDS-PAGE)
3.12 西方墨點法 (Western blotting )
3.13 球狀原生質體(spheroplast)的萃取
3.14 掃描式電子顯微鏡技術 (Scanning Electron Microscopy ;SEM)
3.15 取細菌表面增強拉曼散射光譜
3.16 拉曼光譜分析方法
第四章、結果
第五章、討論
第六章、圖表
參考文獻
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