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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫(Kai-Wun Yeh) | |
| dc.contributor.author | Yih-Feng Hsieh | en |
| dc.contributor.author | 謝懿風 | zh_TW |
| dc.date.accessioned | 2021-06-12T17:54:14Z | - |
| dc.date.available | 2008-02-18 | |
| dc.date.copyright | 2008-02-18 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-02-04 | |
| dc.identifier.citation | 邱崇益 (2005) 過氧化氫酶家族在文心蘭假球莖內之功能性研究。台大植物科學研究所碩士論文。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27035 | - |
| dc.description.abstract | 本實驗室先前從文心蘭扣減式EST cDNA library中得到九個Class Ⅲ的過氧化氫酶基因，有些成員於不同逆境因子(H2O2 、4°C低溫、ABA) 處理下，會被誘導表現；為了解其調控機制，本研究選殖了出文心蘭過氧化氫酶Ppox3-19之1364bp啟動子(peroxidase promoter)，經分析發現有ABRE、GARE、以及與防禦反應相關之TC-rich repeats等cis-elements，將啟動子後接β-glucuronidase(GUS)報導基因並轉殖至阿拉伯芥，研究於不同處理(IAA、ABA、JA、GA3各100 μM，以及培養基中1 %、2 %、3 %、4 %、5 % 蔗糖)及不同大小啟動子片段啟動報導基因GUS之表現情形。於不同處理下，GUS組織染色結果顯示，報導基因皆會在七天大之阿拉伯芥轉殖株的保衛細胞表現，推測於啟動子上具有保衛細胞專一的cis-elements；另外，GUS螢光活性分析顯示，100 μM 之JA、GA3、ABA三種賀爾蒙會提高GUS表現，可達2至 2.5 nmole MU/min/mg protein。此外，隨著培養基中蔗糖濃度越高，GUS表現量會越多，其中5% 蔗糖的處理，GUS表現可達4.3 nmole MU/min/mg protein。
分析不同5’端長度之啟動子於阿拉伯芥轉殖株之表現，推測啟動子於-1343 bp~ -1291 bp處，應有保衛細胞專一性表現之cis-element；在-1291 bp 至-701 bp 除了具有葉肉細胞表現之cis-element 外，也可能具有增強GUS 表現量之cis-elements。 為了解Ppox3-19與ABA訊息傳遞之關係，將Ppox3-19全長轉殖入阿拉伯芥aba 及abi 突變株中，結果顯示abi1中無GUS表現，而abi2中仍有GUS表現於保衛細胞中，顯示Ppox3-19表現於保衛細胞需要ABI1存在於植物細胞中。 由阿拉伯芥轉殖株之結果，我們認為ABA 訊息傳遞路徑之參與是影響POX 基因啟動子於細胞中表現的重要因子，而POX啟動子的-1343 bp~ -1291 bp是決定保衛細胞專一性表達的重要cis-element。 | zh_TW |
| dc.description.abstract | Previous studies had shown that expression patterns under abiotic stress of nine class III peroxidase genes isolated from Oncidium substractive cDNA library. Here we cloned a 1.3 kb of Oncidium peroxidase promoter, named Ppox3-19 that was fused with a GUS reporter gene. By using transgenic Arabidopsis, the promoter activity was investigated under various concentrations of different hormones and sucrose. The results revealed that 7-day-old seedlings of transgenic Arabidopsis specifically exhibited higher GUS activities in guard cells in the presence of 5 % sucrose and 100 μM different hormones such as JA, GA3, and ABA. To further characterize the Ppox3-19, a series of deletions of the promoter fused with were established, and transformed into wild-type Arabidopsis. The results revealed that transgenic plants with the full-length of the promoter showed higher GUS activities in guard cells, rather than those with deletion constructs. We concluded that Ppox3-19 contains guard cell-specific cis-elements between -1341 bp and -1291 bp. The regulatory relationship between ABA and guard cells was also analyzed by transforming the Ppox3-19::GUS construct into Arabidopsis aba and abi mutants. Accordingly, it was found that GUS activities were detected in guard cells of abi2 but not in abi1, which indicates that Ppox3-19 be involved in downstream of ABI1 signaling. Thus, the promoter activity of the peroxidase pox3-19 by higher concentration (> 100 μM) of ABA, and a guard cell-specific cis-element may be present in the promoter region from -1343 bp to -1291bp. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-12T17:54:14Z (GMT). No. of bitstreams: 1 ntu-97-R94b42007-1.pdf: 2038710 bytes, checksum: cc874fb612355eef458dc2f1a8a946c5 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 目錄
口試委員審定書……………………………i 致謝…………………………………………ii 目錄…………………………………………iii 中文摘要……………………………………v 英文摘要……………………………………vi 第一章 前言 第一節 植物防禦機制 …………………1 第二節 過氧化氫酶分類及功能 ………5 第三節 過氧化氫酶之啟動子………………9 第四節 論文之研究目的………………10 第二章 材料與方法 一、 實驗材料…………………………12 二、 實驗方法 第一節 文心蘭過氧化氫酶啟動子(Ppox3-19)之釣取…………13 第二節 載體之構築……………………………………………..23 第三節 基因槍法（particle bombardment）轉殖洋蔥表皮細胞 ………………………………………………………27 第四節 農桿菌之轉型與鑑定…………………………………31 第五節 阿拉伯芥之基因轉殖與轉殖株之鑑定………………32 第六節 阿拉伯芥轉殖株中過氧化氫酶啟動子(Ppox3-19)及其他缺失片段之分析……………………………………………35 第三章 結果 第一節 釣取文心蘭過氧化氫酶Ppox3-19 …………39 第二節 POX基因啟動子(Ppox3-19)載體之構築及轉殖洋蔥表皮細胞……………40 第三節 農桿菌之轉型與鑑定…………………………………40 第四節 阿拉伯芥之基因轉殖與轉殖株之鑑定………………40 第五節 阿拉伯芥轉殖株於植物賀爾蒙誘導處理之GUS螢光活性 分析…………………………41 第六節 不同蔗糖濃度對阿拉伯芥轉殖株之GUS螢光活性 分析…………………………42 第七節 阿拉伯芥轉殖株中過氧化氫酶基因啟動子(Ppox3-19)及其他缺失片段之GUS活性分析…………………………42 第八節 Ppox3-19於阿拉伯芥ABA突變株之GUS螢光活性 分析……………………………………………………43 第四章 討論…………………………………………45 參考文獻…………………………………………………50 圖表………………………………………………………70 附錄………………………………………………………92 | |
| dc.language.iso | zh-TW | |
| dc.subject | 過氧化氫酶 | zh_TW |
| dc.subject | 文心蘭 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.subject | peroxidase | en |
| dc.subject | Oncidium | en |
| dc.subject | promoter | en |
| dc.title | 文心蘭南西品系過氧化氫酶啟動子Ppox3-19基因調控之功能分析 | zh_TW |
| dc.title | Functional studies of promoter activities from Oncidium Gower Ramsey peroxidase gene in Arabidopsis thaliana. | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃鵬林(Pung-Ling Huang),吳克強(Keqiang Wu),謝旭亮(Hsu-Liang Hsieh),陳仁治(Jen-Chih Chen) | |
| dc.subject.keyword | 文心蘭,過氧化氫酶,啟動子, | zh_TW |
| dc.subject.keyword | Oncidium,peroxidase,promoter, | en |
| dc.relation.page | 100 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2008-02-04 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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