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標題: | 阿拉伯芥表現水稻超氧歧化?之研究 Expression and Characterization of Rice Superoxide Dismutase in Arabidopsis thaliana |
作者: | Ming-Kuan Chen 陳銘坤 |
出版年 : | 1998 |
學位: | 碩士 |
摘要: | 超氧歧化?(SOD)是一種含有金屬的酵素,它催化了對細胞有傷害性的超氧分子成為氧和過氧化氫。本論文中建構了一個質體其包含了水稻細胞質型銅鋅超氧歧化?(CuZnSOD)基因,而且此基因由CaMV 35S啟動子所啟動。將這個質體利用農桿菌以抽氣轉殖法送入阿拉伯芥,以得到轉殖系。 本論文工作中,已得到11株含水稻CuZnSOD之轉殖株,北方分析證實轉殖株之葉片中有穩定且大量的轉殖基因表現。轉殖株的SOD活性是野生型的2至4倍,其他抗氧化酵素:catalase, peroxidase和ascorbate peroxidase的活性在轉殖株與野生型的葉片中差異不大。電泳分析結果顯示轉殖株比野生型多出兩種移動速率不同的CuZnSOD分子;經由SDS-PAGE,西方分析及分子量分析後推測兩種分子均屬雙元體,分子量約為26-30 kDa;然而,活性較弱的SOD分子可能是缺少Zn離子所致。 在本論文中亦試驗轉殖株與野生型植株在多種逆境處理後,雖然其植株葉片之抗氧化酵素有些變化,但外觀之生長卻無明顯差異。轉殖株與早開花突變種間雜交後代之開花觀察,結果顯示雜交植株含CuZnSOD及三種早開花突變種之雜交後代植株,均可延遲開花2-5天。 Superoxide dismutase (SOD) is one of the metalloproteins that converts the superoxide radicals to molecular oxygen and hydrogen peroxide. Chimeric gene containing a full length cDNA of rice cytosolic CuZn-superoxide dismutase (CuZnSOD) fused to a CaMV 35S promoter was constructed in binary vectors, mobilized into Agrobacterium, and introduced into wild-type plants of Arabidopsis thaliana, Columbia ecotype, by vacuum infiltration method. Eleven CuZnSOD-transgenic plants were generated. Northern blot analysis showed that the high levels of rice CuZnSOD mRNA transcripts were expressed in the leaves of transgenic plants. Total SOD activity in the transgenic plants was enhanced by 2-4 fold. Catalase, peroxidase and ascorbate peroxidase activities did not show significant change in the transgenic plants. Two different forms of extra CuZnSOD activity in the transgenic plants were observed in the PAGE. The molecular weight of both forms was dimeric size about 26-30 kDa estimated by SDS-PAGE, Western analysis and gel filtration chromatography, but the minor one may lack of Zn as cofactors. It seems that expression of CuZnSOD acitvity can not confer more tolerance in the transgenic plants for several abiotic stresses. The cross made between elf mutants and transgenic plants demonstrated the potential role of SOD involved in the initiation of flowers in Arabidopsis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76366 |
全文授權: | 未授權 |
顯示於系所單位: | 植物科學研究所 |
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