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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王亞男 | |
dc.contributor.author | Yung-Chih Chen | en |
dc.contributor.author | 陳勇至 | zh_TW |
dc.date.accessioned | 2021-06-16T16:39:30Z | - |
dc.date.available | 2017-10-12 | |
dc.date.copyright | 2012-10-12 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-09-11 | |
dc.identifier.citation | 參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63402 | - |
dc.description.abstract | 喜樹鹼(camptothecin, CPT)是一種單萜類吲哚生物鹼,並且是一種目前已廣泛應用在臨床上的抗癌物質,目前發現在喜樹(Camptotheca acuminata)、白花蛇根草(Ophiorrhiza pumila)、青脆枝(Nothapodytes nimmoniana)等植物都具有喜樹鹼,而在青脆枝根部樹皮則具有最高含量的喜樹鹼,但青脆枝中喜樹鹼的生合成卻仍未被完全解析,故本研究從青脆枝當中選殖出四個喜樹鹼生合成相關基因,anthranilate synthase (NnASA)、tryptophan decarboxylase (NnTDC)、cytochrome P450 reductase (NnCPR)、secologanin synthase(NnSLS)及一個拓樸異構酶I(topoisomerase I,NnTop1)。利用離葉系統進行不同的處理研究,當以25 oC以上的高溫或施加1 μM BA的處理,皆可使四個喜樹鹼生合成基因表現量提高;當施加200 μM甲基茉莉酸(MeJA)處理,NnASA、NnTDC及NnCPR表現量會受到誘導,但NnSLS則不受其誘導;而高濃度的氯化鈉溶液處理則會抑制NnASA、NnTDC及NnCPR基因的表現,亦會造成喜樹鹼的累積銳減。
本研究同時發現青脆枝的拓樸異構酶I在喜樹鹼的作用區域中出現兩個胺基酸位置突變(Asn421Lys, Leu530Ile),其可能與青脆枝的抵抗喜樹鹼機制有關。 本研究結果除了可提供青脆枝生產喜樹鹼的最適生長條件之外,亦可作為日後研究喜樹鹼與拓樸異構酶I交互作用的參考依據。 | zh_TW |
dc.description.abstract | Camptothecin (CPT), a monoterpene indole alkaloid, is an important anticancer compound that has been obtained in several plants including Camptotheca acuminata, Ophiorrhiza pumila and Nothapodytes nimmoniana. Current report revealed that the highest level of CPT was detected in root bark of N. nimmoniana, but the CPT biosynthesis in N. nimmoniana is still unclear. In this study, four CPT biosynthetic related genes, including anthranilate synthase (NnASA), tryptophan decarboxylase (NnTDC),cytochrome P450 reductase (NnCPR), secologanin synthase (NnSLS) and a topoisomerase I were cloned from N. nimmoniana. The expression profiling of CPT-related genes were monitored under several treatments for disclosing the pivotal regulatory network. Four CPT - related genes were all induced when plant grew under the warmer ambient temperature condition (> 25 oC). 6-benzyladenine (BA), a kind of cytokinin, is active plant growth regulator to elevate the expression level of all CPT - related genes. NnASA、NnTDC and NnCPR but not NnSLS were induced by 200 μM methyl jasmonate but reduced by high concentration of sodium chloride (salt). Moreover, the CPT amount in leaf treated with high concentration of salt also decreased significantly. Two mutation sites (Asn421Lys, Leu530Ile) existing in topoisomerase I of N. nimmoniana were identified. This result suggested that the CPT-resistant mechanism in N. nimmoniana.
Our results revealed the CPT-biosynthesis and resistance through genetic and molecular network and provided a perspective for cultivating N. nimmoniana. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T16:39:30Z (GMT). No. of bitstreams: 1 ntu-101-R97625060-1.pdf: 3609275 bytes, checksum: ab91370d1b548dc4a6ed4df1b38b5260 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 目錄
口試委員會審定書……………………………………………………………………I 致謝……………………………………….…………………………………………..II 目錄………………………………………………………………….……………….III 表目錄……………………………………………………..……………...………….IV 圖目錄…………………………………………………………………......………….V 中文摘要………………………………………………………….…..……...……..VII 英文摘要………………………………………………………...….…..………….VIII 第一章 前言……………………………………..………………………………..…..1 第一節 植物二次代謝物與喜樹鹼………..……………………………..……..1 第二節 青脆枝之生物特性與應用…………..………………………..………..4 第三節 喜樹鹼的藥性機制……………………..…………………..…………..6 第四節 植物喜樹鹼的生合成路徑………………………………..……………8 第五節 本論文研究方向………………………….……………..…………….10 第二章 材料與方法…………………………………………………..……………..11 第一節 喜樹鹼生合成關鍵基因選殖………………………....………………11 第二節 喜樹鹼生合成基因於青脆枝基因體之套數……..…….…………….21 第三節 基因表現量分析…………………………………..………..…………27 第四節 喜樹鹼含量分析..………………………………..……………...…….29 第三章 結果與討論…………………………………………..……….…………….31 第一節 青脆枝拓樸異構酶I與喜樹鹼合成基因的選殖及其基因套數分析31 第二節 青脆枝中喜樹鹼生合成相關基因表現量分析……………………....34 第三節 喜樹鹼含量分析………………………………………...…………….38 第四章 結論…………………………………………………..……….…….………40 參考文獻………………………………………………………….………………….42 附表…………………………………………………………………………….…….49 附圖…………………………………………………………………….…………….54 | |
dc.language.iso | zh-TW | |
dc.title | 青脆枝中喜樹鹼生合成基因的表現與其拓樸異構酶I的胺基酸序列分析 | zh_TW |
dc.title | Gene expression of Camptothecin-biosynthetic genes and Sequence analysis of Camptothecin-resistant Topoisomerase I in Nothapodytes nimmoniana | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-1 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 葉開溫 | |
dc.contributor.oralexamcommittee | 林敏宜,蕭英倫,李明仁 | |
dc.subject.keyword | 青脆枝,喜樹鹼生合成基因,喜樹鹼,拓樸異構?l, | zh_TW |
dc.subject.keyword | Nothapodytes nimmoniana,camptothecin biosynthetic genes,camptothecin,topoisomerase I, | en |
dc.relation.page | 87 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-09-13 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 森林環境暨資源學研究所 | zh_TW |
顯示於系所單位: | 森林環境暨資源學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-101-1.pdf 目前未授權公開取用 | 3.52 MB | Adobe PDF |
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