Taipei Medical University Institutional Repository:Item 987654321/5056
English  |  正體中文  |  简体中文  |  全文筆數/總筆數 : 45073/58249 (77%)
造訪人次 : 2390045      線上人數 : 161
RC Version 7.0 © Powered By DSPACE, MIT. Enhanced by NTU Library IR team.
搜尋範圍 查詢小技巧：
  • 您可在西文檢索詞彙前後加上"雙引號"，以獲取較精準的檢索結果
  • 若欲以作者姓名搜尋，建議至進階搜尋限定作者欄位，可獲得較完整資料
    •  進階搜尋


    請使用永久網址來引用或連結此文件: http://libir.tmu.edu.tw/handle/987654321/5056


    題名: 以蛋白質體學法分析短鏈脂肪酸代謝異常ENU突變鼠
    Proteomic analysis of ENU mice with abnormal defect of short chain fatty acid
    作者: 黃小珍
    Hsiao-Chen Huang
    貢獻者: 保健營養學研究所
    關鍵詞: 致突變劑
    串聯質譜儀
    短鏈脂肪酸
    二維電泳法
    介質輔助雷射脫附游離-飛行式質譜儀
    ethyl-nitrosourea
    MS/MS tandem mass
    C4-OH
    Two-dimensional electrophoresis
    MALDI-TOF
    日期: 2006
    上傳時間: 2009-09-10 18:02:54 (UTC+8)
    摘要: 中文摘要
    本研究目的為利用二維電泳法 (Two-dimensional electrophoresis ; 2-DE) 以找出ENU (ethyl-nitrosourea)突變小鼠表現差異的蛋白質點並進行質譜分析，以瞭解突變小鼠短鏈脂肪酸代謝異常原因，並且希望能應用此一模式動物於脂肪酸的代謝研究及探討人類相關臨床疾病。我們利用ENU注射動物方法藉以誘發大量基因突變小鼠的產生，經由繁殖配對至第三代後，再以串聯式質譜儀篩得血漿短鏈脂肪酸 (C4-OH) 代謝異常突變小鼠。實驗小鼠 (C57BL/6J) 分為正常小鼠及血漿C4-OH含量較正常小鼠高4倍標準差的ENU突變小鼠，兩組小鼠分別犧牲後取其肝臟、肌肉組織，進行粒線體蛋白質萃取後以SDS-PAGE進行一維電泳，然而突變小鼠和正常小鼠粒線體蛋白質表現量並無差異。因此，更進一步利用二維電泳法進行突變小鼠和正常小鼠粒線體蛋白質表現的比較。研究結果顯示突變小鼠和正常小鼠的肌肉粒線體存有差異的蛋白質點，經由介質輔助雷射脫附游離-飛行式質譜儀 (Matrix-assisted laser desorption / ionization time-of-flight mass spectrometry; MALDI-TOF) 鑑定出差異蛋白質點的身份。在突變小鼠表現量增加的蛋白質為磷酸化的myosin regulatory light chain 2、tropomysin 1 α chain、 myosin light chain 1，表現量減少的蛋白質為非磷酸化的myosin regulatory light chain 2、calsequestrin-1 precursor、adenylate kinase isoenzyme 1、ATP synthase D chain。然而這些蛋白質表現的消長是否與突變鼠體內C4-OH含量上升有關，仍有待更進一步的研究加以驗證。
    關鍵詞：致突變劑 (ethyl-nitrosourea; ENU)、串聯質譜儀、短鏈脂肪酸、二維電泳法(Two-dimensional electrophoresis ; 2-DE)、介質輔助雷射脫附游離-飛行式質譜儀(Matrix-assisted laser desorption / ionization time-of-flight mass Spectrometry; MALDI-TOF)
    Abstract
    The purpose of this study was to develop a 2-DE coupling with MALDI-TOF analysis to find the differential proteins in ENU mouse which could explore the cause of genetic defect of short chain fatty acid in mice. We hope that the C4-OH animal model can provide us to investigate fatty acid metabolism related to human genetic disease. We generated ENU- mutagenized mouse and applied to MS/MS tandem mass spectrometry to screen third generation progeny of ENU-mutagenized mouse for abnormalities in the pathway of fatty acid metabolism. The short chain fatty acid (C4-OH) level in ENU mice was three to four folds than normal mice. After protein evaluation, there was no differential protein in mitochondria fraction between normal and ENU mice by SDS-PAGE. Surprisingly, there were eleven differential proteins between normal and ENU mice in muscle in 2-DE approach. These protein spots were applied to MALDI-TOF for futher identification. In results, ENU mouse showed higher protein expressions in phosphorylated myosin regulatory light chain 2, myosin light chain 1 and tropomyosin 1αchain and lower expressions in myosin regulatory light chain 2, adenylate kinase isoenzyme, ATP synthase D chain and calsequestrin-1 precursor in ENU mouse to normal control. These differential proteins expression whether involve to increase C4-OH value in mice needs further study to verify.
    Key words：Two-dimensional electrophoresis, ethyl-nitrosourea, C4-OH, MS/MS tandem mass, MALDI-TOF
    資料類型: thesis
    顯示於類別:[保健營養學系暨研究所] 博碩士論文

    文件中的檔案:

    檔案 描述 大小格式瀏覽次數
    摘要.doc28KbMicrosoft Word107檢視/開啟
    摘要.pdf117KbAdobe PDF128檢視/開啟
    摘要.ppt125KbMicrosoft Powerpoint163檢視/開啟
    摘要.ps630KbPostscript68檢視/開啟


    在TMUIR中所有的資料項目都受到原著作權保護.

    TAIR相關文章

    著作權聲明 Copyright Notice

    • 本平台之數位內容為臺北醫學大學所收錄之機構典藏，包含體系內各式學術著作及學術產出。秉持開放取用的精神，提供使用者進行資料檢索、下載與取用，惟仍請適度、合理地於合法範圍內使用本平台之內容，以尊重著作權人之權益。商業上之利用，請先取得著作權人之授權。

      The digital content on this platform is part of the Taipei Medical University Institutional Repository, featuring various academic works and outputs from the institution. It offers free access to academic research and public education for non-commercial use. Please use the content appropriately and within legal boundaries to respect copyright owners' rights. For commercial use, please obtain prior authorization from the copyright owner.

    • 瀏覽或使用本平台，視同使用者已完全接受並瞭解聲明中所有規範、中華民國相關法規、一切國際網路規定及使用慣例，並不得為任何不法目的使用TMUIR。

      By utilising the platform, users are deemed to have fully accepted and understood all the regulations set out in the statement, relevant laws of the Republic of China, all international internet regulations, and usage conventions. Furthermore, users must not use TMUIR for any illegal purposes.

    • 本平台盡力防止侵害著作權人之權益。若發現本平台之數位內容有侵害著作權人權益情事者，煩請權利人通知本平台維護人員([email protected])，將立即採取移除該數位著作等補救措施。

      TMUIR is made to protect the interests of copyright owners. If you believe that any material on the website infringes copyright, please contact our staff([email protected]). We will remove the work from the repository.

    Back to Top
    DSpace Software Copyright © 2002-2004  MIT &  Hewlett-Packard  /   Enhanced by   NTU Library IR team Copyright ©   - 回饋