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標題: | 水稻銅鋅超氧歧化?在阿拉伯芥轉殖株葉中其蛋白質衰變之研究 Turnover of Rice CuZnSOD in the Leaves of SOD-Overexpressing Arabidopsis |
作者: | Yen-Ju Chen 陳晏如 |
出版年 : | 1998 |
學位: | 碩士 |
摘要: | 以大量表現水稻銅鋅超氧歧化?(CuZnSOD)之阿拉伯芥轉殖株和野生株為材料,利用單向電泳法來分析野生株和轉殖株之蛋白質,發現除了轉殖基因產物水稻CuZnSOD之外,兩者之蛋白質含量及種類大致相同。以雙向電泳法分析其蛋白質及西方點墨分析,則發現轉殖株至少多了5個pI值不同但分子量相同可被水稻CuZnSOD抗體所辨認之額外蛋白質。取阿拉伯芥之葉柄進行35S-methionine活體標定之實驗,結果顯示野生株及轉殖株對於35S-methionine皆有21-29%的吸收率,約有1?1.5%進入新合成的蛋白質。萃取之蛋白質利用電泳分析和自動放射顯影技術,分析其標定期間新合成之蛋白質,轉殖株中大部份蛋白質的衰變與野生株相似,而轉殖基因表現之CuZnSOD其衰變速度較大部份蛋白質為快。若以單向電泳分析則水稻之CuZnSOD於轉殖株葉片中約20小時之內可衰變50%的量。本論文並討論以35S-methionine活體標定葉片後再以雙向電泳分析CuZnSOD之衰變結果。 In order to estimate the turnover of CuZn-superoxide dismutase (CuZnSOD) and the gene products in transgenic Arabidopsis, protein patterns of SOD-overexpressor and wild-type plants from Arabidopsis Columbia ecotype were analyzed by 1 D-PAGE or 2D-PAGE. After the pulse and chase experiment with 35S-methionine was done in the shoot parts of plants, the soluble polypeptides from the leaves were extracted. The de novo polypeptides were analyzed with one- or two-dimensional gel electrophoresis and autoradiography. The uptake of 35S-methionine and incorporation into polypeptides in the transgenic Arabidopsis were about 21?29% and 1.5%, respectively, which were similar to those observed in the wild-type plants. The turnover of rice CuZnSOD in SOD-transgenic Arabidopsis was estimated to be within 20 hr by the 1D-PAGE and autoradiogram image analysis. 2D-PAGE and autoradiogram image analysis were also used and discussed in the turnover study of CuZnSOD in SOD-transgenic Arabidopsis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76379 |
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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