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頁籤選單縮合
| 題 名 | 應用可誘導轉位子移除轉基因植物之篩選標記=Using Transposons to Remove Selectable Marker from Transgenic Plants |
|---|---|
| 作 者 | 常玉強; 謝嘉珊; | 書刊名 | 中華農學會報 |
| 卷 期 | 7:1 民95.02 |
| 頁 次 | 頁43-54 |
| 分類號 | 434.28 |
| 關鍵詞 | 轉基因作物; 可誘導轉位子; 篩選標記; 標的基因; Transgenic plants; Inducible transposon; Selectable marker gene; Target gene; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 執行轉基因作物時,除了將轉殖標的基因外,為了篩選成功之轉基因植物,往往必須同時轉殖篩選標記基因。目前所使用之篩選標記基因大多為NPTⅡ、HPTⅡ、bar基因,該等基因表現後可耐殺草劑或抗生素。然而消費者質疑篩選標記基因可能漂移至其他非轉基因品種,因此,獲得轉基因作物後,保留標的基因並移除篩選標記基因為推廣轉基因作物之重要工作。先前,作者曾建構--「可誘導轉位子」--將轉位基因建構於可受水楊酸誘導之PR-1a啟動子後—轉位現象可受水楊酸而控制。據此,本研究將抗Hygromycin之HPT基因作為篩選標記基因建構於「可誘導轉位子」內,並分別轉殖入番茄及水稻。執行轉基因操作時,歸選轉基因植株時不誘導轉位子而可有效執行篩選工作,待篩選出成功轉基因作物後,施加水楊酸使含篩選標記基因之轉位子跳至其他染色體。上述「可誘導轉位子」並建構於1’啟動子與螢火蟲之冷光基因中,「可誘導轉位子」轉位後冷光基因才表現,本研究以番茄轉殖株為材料偵測出具冷光活性之番茄轉殖株,並以南方氏墨點分析證明轉位子跳離,該轉基因番茄自交後將獲得轉基因作物僅含冷光基因而不抗Hygromycin之植株。此外,利用TAIL-PCR技術及水稻定序資料庫,本究究證明含上述構築之水稻T-DNA及已跳離轉位子分別位於不同染色體。 |
| 英文摘要 | The use of transgenic technology for agricultural purposes has encountered several challenges. One concern is related to the presence in transgenic crop plants of market genes conferring antibiotic or herbicide resistance. Although no scientific basis has been determined for these concerns, removal of marker genes would likely hasten the public acceptance of transgenic crops. To this end, in the present study, an inducible transposon system has been constructed to overcome these limitations. A fusion of the PR-1a promoter with the Ac transposase gene was inserted together with a hygromycin resistance gene (selectable marker) between 5’ and 3’ end of the Ac transposon, yielding an inducible transposable element. The element was constructed between the 1’ promoter and the luciferase gene (target gene) from firefly. These constructs were introduced into tomato plants. After induction by salicylic acid, the transposons in transgenic tomato transpose resulted in the luciferase light emission. The progeny of these tomato plants were analyzed by luciferase verses hygromycin resistance activity as well as Southern blot analysis. the results indicate that the inducible transposon transposed into new positions which were unlinked to the T-DNA locus, demonstrating that the marker-free transgenic progeny harboring the target (luciferase) gene was obtained. Additionally, the same strategy was performed in rice plants, in an attempt to yield marker-free transgenic rice. TAIL-PCR analysis of the flanking sequences of T-DNA and transposon indicated that the transposons can be induced to transpose other chromosome locus from the T-DNA in transgenic rice plants. |
本系統中英文摘要資訊取自各篇刊載內容。