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    標題: 利用多重PCR技術檢測基因改造玉米
    Detection of genetically modified maize by multiplex polymerase chain reaction
    作者: 戴永萍
    Yung-ping Tai
    貢獻者: 徐阿里
    張竣凱
    嘉南藥理科技大學:生物科技研究所
    關鍵字: 即時聚合酶鏈鎖反應
    聚合酶鏈鎖反應
    基因改造玉米
    基因改造作物
    多重聚合酶鏈鎖反應
    Polymerase chain reaction ( PCR )
    Genetically modified plants
    Genetically modified maize
    Multiplex PCR
    Real-time polymerase chain reaction (real-time)
    日期: 2008
    上傳時間: 2008-12-29 15:23:53 (UTC+8)
    摘要: 基因改造作物在近年來被廣泛種植並應用於農業生技產品技術的發展,且隨著愈來愈多基因改造作物的商品市場化並大規模生產,引起基因食物的爭論疑慮,例如基因食品的安全性、對生態環境的影響和倫理道德等問題。因此我們以標的基因設計 NK603、Event176、T25、GA21、MON863、MON810、TC1507、Bt11 及內部對照基因 Zein 之專一性引子組,建立基因改造玉米之聚合酶鏈鎖反應 ( PCR ) 定性檢測方法,並利用多重聚合酶鏈鎖反應 ( Multiplex PCR ) 同時檢測這8種基因改造玉米品項( event ),再進行電泳分析得到特定的 DNA 片段檢測結果。本實驗結果顯示利用多重聚合酶鏈鎖反應的方法同時置入多對引子組,只需使用一個微量離心管,透過一個檢驗步驟,即可有效精準鑑別篩選基因改造玉米多種疊加基因品項。此法不僅可減少繁複的操作程序、降低操作時的錯誤機率,更可節省時間及成本。再輔以即時聚合酶鏈鎖反應 ( real-time PCR ) 檢測基因改造玉米含量,將有助於建立基因改造玉米產品標示管理。
    Biotechnology has been widely used in modern agriculture and related industries. Hundreds of genetically modified ( GM ) plants have been approved for commercial production. Several controversial issues such as food safety, environment risk, and ethical concerns are being discussed. The purpose of this study was to develop multiplex polymerase chain reaction ( multi-PCR ) method for the simultaneous detection of eight events in genetically modified ( GM ) maize. The eight primer pairs were constructed specifically for eight respective GM events of Bt11, Event176, GA21, MON810, MON863, NK603, T25, and TC1507 and a primer pair for an endogenous reference gene, Zein. Its amplified products could be distinguished by agarose gel and capillary electrophoreses based on their different lengths. This method can specifically detect all eight events in one-tube with high sensitivity. The results indicate that this multiplex PCR method could be used for the identification and detection of GM maize. Furthermore, this method could be used for the event-specific monitoring on hybrid stack progenies between two different events of GM maize. The established event-specific real-time PCR detection systems in this study are suitable for the identification and quantification of these GM maizes.
    關聯: 校內校外均不公開
    顯示於類別:[生物科技系(所)] 博碩士論文

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