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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 常玉強(Yuh-Chyang Charng) | |
| dc.contributor.author | Hsiu-Chun Yang | en |
| dc.contributor.author | 楊琇淳 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:08:02Z | - |
| dc.date.available | 2013-02-24 | |
| dc.date.copyright | 2010-02-24 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-02-05 | |
| dc.identifier.citation | Andres, D.A., Dickerson, I.M., Dixon, J.E. (1990) Variants of the carboxyl-terminal KDEL sequence direct intracellular retention, J Biol Chem, 265, 5952–5955.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45187 | - |
| dc.description.abstract | 本試驗使用持續表現和水楊酸誘導表現系統，鑑別來自水稻的OsCPs (Os09g21370) 基因之功能。OsCPs基因產物為半胱胺酸蛋白酶。藉由前人PCR反應修飾OsCPs基因，可產生N端缺失30和60個胺基酸，研究三種結構之半胱胺酸酶是否可因持續表現於細胞質而降解蛋白質使細胞致死。因此本試驗使用持續表現CaMV-35S啟動子表現全長，N端缺失30個或缺失60個胺基酸的OsCPs轉殖阿拉伯芥、菸草和水稻，偵測轉殖株基因表現和性狀分析。此外也使用水楊酸誘導之PR-1a啟動子表現全長OsCPs轉殖菸草，偵測轉殖株經水楊酸誘導後OsCPs基因表現和性狀分析。
CaMV-35S啟動子之持續表現系統中，阿拉伯芥、菸草和水稻皆成功獲得轉殖株，RT-PCR也偵測轉殖株外源基因皆有表現，但並無致死性狀。此系統試驗結果顯示，持續表現全長和缺失的OsCPs基因對水稻、菸草和阿拉伯芥轉殖效率有些微影響，但轉殖株仍可持續生長至成株。結果顯示，全長和缺失的OsCPs基因表現皆無立即致死性。另外，PR-1a表現系統表現全長OsCPs基因的轉殖株，經誘導劑誘導後，確實可偵測OsCPs基因表現，且植株並無致死性狀。再次證實OsCPs基因無致死性。論文最後討論本試驗未來的發展方向。 | zh_TW |
| dc.description.abstract | The OsCPs (Os09g21370) gene from rice (Oryza sativa ssp japonica cv. Nipponbare) is expected to encode a cysteine protease, which has potential uses as cytotoxins for cell death when it triggered by constitutive promoter. Previously, three forms of this gene have been cloned as full-length and truncated (30 or 60 amino acids deleted from N-terminal). In this work, to determine and characterize the OsCPs gene’s functions, three forms of this gene were expressed constitutively or conditionally, by ligating the genes independently with CaMV-35S or PR-1a inducible promoter. The resulted six constructs were introduced into Arabidopsis, tobacco and rice plants by Agrobacterium-mediated transformation system. Analysis of the transgenic plants containing full-length and truncated OsCPs gene triggered by the CaMV-35S promoter indicate that neither full-length nor truncated form of the OsCPs gene leads to cell death. The expression of each construct was confirmed by RT-PCR technique with the mRNA templates extracted from the corresponding transgenic plants. Also, salicylic acid was applied to induce transgenic tobacco plants containing OsCPs gene triggered by the PR-1a promoter. Again, no lethal effect was observed and RT-PCR experiments demonstrated the expression of each form of the gene. Several ways were discussed to resolve the expression problems of OsCPs gene and the availability of the inducible system in the future. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:08:02Z (GMT). No. of bitstreams: 1 ntu-99-R96621102-1.pdf: 5171579 bytes, checksum: 42db2dfc1330cb40aea00a59fe5ef77e (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 誌謝................................................. i
中文摘要............................................. ii 英文摘要............................................. iii 目錄................................................. iv 圖目錄............................................... v 表目錄............................................... vi 附錄目錄............................................. vii 縮寫字中英對照表..................................... viii 壹、前言............................................. 01 貳、前人研究......................................... 02 参、試驗設計與流程................................... 12 肆、材料與方法....................................... 14 伍、結果............................................. 27 陸、討論............................................. 38 柒、未來展望......................................... 43 捌、參考文獻......................................... 44 表................................................... 50 圖................................................... 56 附錄................................................. 76 | |
| dc.language.iso | zh-TW | |
| dc.subject | 水楊酸誘導啟動子 | zh_TW |
| dc.subject | 半胱胺酸蛋白酶 | zh_TW |
| dc.subject | 細胞程序性死亡 | zh_TW |
| dc.subject | 致死基因 | zh_TW |
| dc.subject | PCD | en |
| dc.subject | Cysteine protease | en |
| dc.subject | PR-1a promoter | en |
| dc.subject | lethal gene | en |
| dc.title | 水稻半胱胺酸蛋白酶功能之研究 | zh_TW |
| dc.title | Function Studies of a Rice Cysteine Protease Gene | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 杜鎮(Jenn Tu),洪傳揚(Chwan-Yang Hong) | |
| dc.subject.keyword | 半胱胺酸蛋白酶,細胞程序性死亡,致死基因,水楊酸誘導啟動子, | zh_TW |
| dc.subject.keyword | Cysteine protease,PCD,lethal gene,PR-1a promoter, | en |
| dc.relation.page | 86 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-02-05 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 農藝學研究所 | zh_TW |
| 顯示於系所單位： | 農藝學系 | |
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