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油膜蛋白基因轉移至甘藍之研究

By 劉程煒, 林榕華, 曾夢蛟, Cheng-Wei Liu, Rong-Hwa Lin and Menq-Jiau Tseng

Abstract

油膜蛋白基因(oleosin)為富含於種子特殊胞器-油體(oilbody)表面的蛋白,其功能為穩定油體,增加油體與磷脂酵素的接觸面積,與三酸甘油脂的合成與分解有關。本試驗將分離自水稻油膜蛋白基因(OLE),構築到以rubisco small subunit(rbc S)及CaMV 35S啟動子的轉殖載體,並利用農桿菌基因轉移的方式,將帶有目標基因的pKcOn、pKrOn質體分別送入甘藍(新豐、初秋、高峰)的子葉或下胚軸中。試驗結果顯示,經農桿菌感染供試之三個品種的甘藍下胚軸之轉殖再生率介於2.7%-7.7%之間,子葉的轉殖再生率均為零。再生植株經PCR、南方slot轉漬雜交做初步篩選後,再以南方電泳墨點雜交法分析檢驗,均可轉殖植株的DNA上偵測到雜交訊號。北方雜交墨點分析的結果顯示,存在轉殖植株體內的油膜蛋白基因皆可轉錄出RNA。轉殖攜帶有bc S啟動子之油膜蛋白基因的再生植株,其基因的表現以CaMV 35S為啟動子者為佳。Oleosins are a group of Mr 15`26 kD amphipathic proteins which are associated with the surface of oil bodies in lipid-storing tissues such as seeds and pollen. The role of oleosin in oil body is biogenesis and structural stabilization. This research focuses on the use of cabbage (Brassica oleracea L. var. capitata L.) as a model system to establish the gene transfer technology, and possibility for improvement of cabbage with nutrient quality, through the art of genetic engineering.\ud Oleosin (OLE) gene was isolated from rice constructed into plant transformation vectors driven by CaMV 35S or rubisco small subunit (rbc S) promoter, and introduced into the hypocotyl and cotyledon of cabbage Agrobacterium-mediated transformation. Regenerated plants of cabbage wereobtained after transformation. The regeneration rate for cabbage was among 2.7% to 7.7%. Only few plants were regenerated from the cotyledon of cabbage. The regenerated plants were examined by PCR, Southern, and Northern hybridization. The results indicated that the expression of constructed genes was higher in transgenic plants driven by the rbc S promoter than CaMV 35S promoter

Topics: 基因轉移, 甘藍, 油膜蛋白基因, Cabbage, Gene transfer, Oleosin
Year: 2014
OAI identifier: oai:ir.lib.nchu.edu.tw:11455/81541
Journal:

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