應用自由基誘導物2,2’-Azobis (2-amidinopropane) hydrochloride (AAPH)模擬氧化逆境：研究大豆之抗氧化酵素Superoxide dismutase (SOD)的基因表現

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巫盈忠(Ying-Chung Wu) 查詢紙本館藏

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應用自由基誘導物2,2’-Azobis (2-amidinopropane) hydrochloride (AAPH)模擬氧化逆境：研究大豆之抗氧化酵素Superoxide dismutase (SOD)的基因表現
(Molecular studies of Superoxide dismutase (SOD) in soybean by a free radical initiator AAPH)

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摘要(中)

在本研究中，使用一水溶性的自由基誘導物 [2,2’’-Azobis (2-amidinopropane) hydrochloride]（AAPH）來誘發逆境氧化的產生，然後探討逆境生成時大豆在基因 (sod) 表現的變化。應用cytosolic Cu/Zn sod基因的探針來探討基因的表現情形，然後再測定超氧歧化?活性和活性電泳來了解其間的相關性。
從本研究之結果得知cytosolic Cu/Zn sod基因的表現會受AAPH刺激後而降低，但其活性會有升高的情況。根據1996年Pitcher和Zilinskas的報導指出cytosolic Cu/Zn sod對臭氧有較明顯的變化，從本試驗結果指示AAPH可能可以當作一誘發環境逆境的有利研究之工具，來替代如臭氧及其他環境因子壓力的實驗來模擬逆境之研究。

摘要(英)

In this study, a water soluble free radical initiator [2,2’’-azobis (2-amidinopropane) dihydrochloride ] AAPH was used as a research tool to induce stress, and to investigate the gene expression in soybean. Using cytosolic Cu/Zn sod gene probe showed AAPH can affect the sod gene expression and SOD activity in soybean leaves. Results showed the level of the sod transcripts decreased in response to AAPH, but the level of SOD activity increased. Followed the Pitcher and Zilinskas report in 1996, the cytosolic Cu/Zn sod gene and SOD were also responded to ozone stress. From this result suggests that AAPH could be as a research tool to study the environmental stress, to replace an expensive ozone facilities.

關鍵字(中)

★ 活性電泳
★ 自由基
★ 活性態氧
★ 超氧歧化

關鍵字(英)

★ free radical
★ AOS
★ active oxygen species
★ SOD
★ superoxide dismutase
★ native PAGE

論文目次

1. 中文摘要……………………………………………………………Ⅱ
2. Abstract……………………………………………………………...Ⅲ
3. 第一章前言…………………………………………………………1
4. 第二章材料與方法…………………………………………..……..7
5. 第三章結果………………………………………………….…….19
6. 第四章討論………………………………………………………..22
7. 圖一、RNA轉印裝置………………………………………………25
圖二、以AAPH誘導自由基產生對大豆病理變化………………26
圖三、Cytosolic Cu/Zn sod 基因PCR產物………………….…….27
圖四、大豆cytosolic Cu/Zn sod 基因DNA序列之分析…………28
圖五、以DIG-Labeling合成cytosolic Cu/Zn sod探針…………….29
圖六、以1﹪formaldehyde gel鑑定總核醣核酸抽取品質….……..30
圖七、北方轉印法實驗結果………………………………………..31
圖八、超氧歧化?活性測定………………………………………..32
圖九、超氧歧化?活性電泳………………………………………..33
8. 參考文獻……………………………………………………………34

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指導教授

李賢淇

審核日期

2000-7-17

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