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標題: Studies on Transformation of Green Fluorescent Protein Gene (egfp) into the Rice
作者: 陳俊麟
Jiun-Lin Chen
Ming-Kun Ji
Menq-Jiau Tseng
關鍵字: 基因轉殖;Gene transformation;綠色螢光蛋白基因;水稻;Rice;Green fluorescent protein (GFP)
Project: Horticulture NCHU, Volume 31, Issue 1, Page(s) 55-68.
興大園藝, Volume 31, Issue 1, Page(s) 55-68.
In this study, green fuorescent protein gene (egfp) of jelly fsh was transformed into the rice (Oryza sativa L. cv. Tainung 67) via Agrobacterium-mediated transformation to explore the possibility of using egfp genes as selection marker in gene transformation. The regenerated percentage of G418 resistance rice calli was 7 to 12.5%. The transformed plants were fertile and normal in appearance. A 750 bp fragment of egfp could be amplified from transformed rice plants by PCR analysis. Emission of green fluorescence could be observed from egfp transformed rice calli and regenerated plants after excitation with blue light. In roots, high emission of green fluorescence was found in the pKcEGFP (CaMV35S promoter) transformed plants. In leaves and spikes, higher emission of green fluorescence was found in the pKrRGFP (rbcS promoter) transformed plants than those of pKcEGFP transformed plants. The data show the feasibility of using EGFP as a convenient and rapid reporter to select for the genetically modified rice plantlets both in vivo and in vitro.

本研究利用農桿菌轉殖法將綠色螢光蛋白基因(egfp)轉殖到'台農度67號'水稻(Oryza sativa,探討以egfp基因作為篩選標誌基因的可能性。轉殖之水稻癒傷組織經2-3週後,於螢光顯微鏡檢測即可發現綠色螢光,並可誘導再生植株,轉殖再生率為7-12.5%。再生之轉殖植株生長正常,以PCR檢測綠色螢光蛋白基因(egfp)的結果,顯示所有檢測的植株皆具有75Obp的egfp吩基因片段。轉殖植株經照設藍光後,可發散出綠色螢光。在根部的組織,以轉殖pKcEGFP(CaMV35S啟動子)的轉殖株表現最佳,可觀察到最亮的綠色螢光。葉片與稻穗的部份則以轉殖pKrEGFP(rbcS啟動子)的轉殖株表現較佳。試驗結果顯示,可利用egfp基因在水稻基因轉殖過程中,作為早期篩選之用。
ISSN: 0255-5921
Appears in Collections:第31卷 第01期

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