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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 曲芳華(Fang-Hua Chu) | |
| dc.contributor.author | Wei-Lin Fan | en |
| dc.contributor.author | 范瑋琳 | zh_TW |
| dc.date.accessioned | 2022-11-25T05:36:38Z | - |
| dc.date.available | 2027-02-10 | |
| dc.date.copyright | 2022-02-18 | |
| dc.date.issued | 2022 | |
| dc.date.submitted | 2022-02-10 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82136 | - |
| dc.description.abstract | 萜類化合物屬於植物的一種特殊代謝物，除了對於植物適應環境有重要作用，因其所具有生物活性常被運用於醫療保健用途。食茱萸乃芸香科花椒屬植物，分布於臺灣中低海拔，其葉子具特殊香味，原住民常以其嫩葉為傳統食材。經前人研究已知食茱萸葉部含有多種揮發性萜類化合物，單萜化合物應為其主要香氣成分。本研究以食茱萸葉轉錄體資料，藉由生物資訊軟體與芸香科中已發表之揮發性萜類合成酶基因序列作參考比對，篩選出五個單萜合成酶候選基因，以聚合酶連鎖反應獲得全長序列，並以酵素體外活性反應試驗與大腸桿菌共表現系統得到產物以便進行後續酵素功能鑑定，結果顯示Za_mTPS1主要產物為sabinene，Za_mTPS2主要產物為geraniol，Za_mTPS3、Za_mTPS4主要產物皆為linalool，Za_mTPS5主要產物則為(E)-β-ocimene。此外，亦以反轉錄聚合酶連鎖反應分析不同葉組織之基因表現量，以及嘗試以轉基因毛狀根培養生產目標化合物。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T05:36:38Z (GMT). No. of bitstreams: 1 U0001-0802202215022800.pdf: 7118271 bytes, checksum: 441d00ab02aefff0c911c5707ef82f80 (MD5) Previous issue date: 2022 | en |
| dc.description.tableofcontents | 誌謝 i 摘要 ii 圖目錄 vi 表目錄 ix 第一章 前言 1 第二章 文獻回顧 3 2.1 萜類化合物基本介紹 3 2.2 萜類化合物生合成途徑 4 2.3 萜類合成酶特性 7 2.4 植物單萜化合物功能 9 2.5 單萜化合物之應用 10 2.6 毛狀根培養 11 2.7 食茱萸活性成分與萜類研究 13 第三章 材料與方法 15 3.1 實驗材料 15 3.2 食茱萸RNA萃取與cDNA合成 15 3.3 食茱萸單萜合成酶基因選殖 15 3.4 親緣關係樹狀圖分析 (phylogenetic tree) 18 3.5 蛋白質表現載體之構築 18 3.6 蛋白質特性預測 19 3.7 重組蛋白質表現 – 西方墨點法 (Western Blot) 20 3.8 重組蛋白質純化 21 3.9 蛋白質體外活性試驗 (in vitro assay) 22 3.10 大腸桿菌共表現系統 (co-expression) 試驗 (in vivo) 23 3.11 目標基因於葉組織之表現量分析 24 3.12 食茱萸葉之揮發性化合物萃取 25 3.13 毛狀根培養 25 3.13.1 試驗植物之無菌苗準備 25 3.13.2 構築根毛農桿菌之目標基因表現載體 25 3.13.3 植物組織感染 27 3.13.4 轉基因毛狀根基因表現確認 28 3.13.5 毛狀根之產物萃取 29 3.14 氣相層析質譜儀分析 29 第四章 結果 31 4.1 單萜合成酶基因選殖 31 4.2 重組蛋白質之表現 36 4.3 重組蛋白質之純化 36 4.4 蛋白質體外活性試驗反應 38 4.5 大腸桿菌共表現系統分析 41 4.6 食茱萸葉組織基因表現量分析 44 4.7 食茱萸葉之揮發性化合物萃取 44 4.8 轉基因毛狀根基因表現確認 46 4.9 轉基因毛狀根之產物分析 47 第五章 討論 54 5.1 食茱萸單萜合成酶基因序列分析 54 5.2 Za_mTPS3、Za_mTPS4功能探討 54 5.3 Sabinene synthase之活性位點探討 58 5.4 Ocimene synthase於被子植物之演化探討 61 5.5 食茱萸葉組織基因表現量分析 63 5.6 食茱萸葉揮發物分析 63 5.7 轉基因毛狀根困境 64 第六章 結論 68 第七章 參考文獻 69 | |
| dc.language.iso | zh-TW | |
| dc.subject | 萜類合成酶 | zh_TW |
| dc.subject | 食茱萸 | zh_TW |
| dc.subject | 植物香氣成分 | zh_TW |
| dc.subject | 單萜類化合物 | zh_TW |
| dc.subject | 毛狀根培養 | zh_TW |
| dc.subject | monoterpenoid | en |
| dc.subject | hairy root | en |
| dc.subject | terpene synthase | en |
| dc.subject | plant aroma compounds | en |
| dc.subject | Zanthoxylum ailanthoides | en |
| dc.title | 食茱萸單萜合成酶基因選殖與功能鑑定 | zh_TW |
| dc.title | Cloning and Characterization of Monoterpene Synthase Genes from Zanthoxylum ailanthoides | en |
| dc.date.schoolyear | 110-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鍾振德(Yi-Cheng Weng),何振隆(Yih-Dao Wu),孫英玄,王升陽 | |
| dc.subject.keyword | 食茱萸,植物香氣成分,萜類合成酶,單萜類化合物,毛狀根培養, | zh_TW |
| dc.subject.keyword | Zanthoxylum ailanthoides,plant aroma compounds,terpene synthase,monoterpenoid,hairy root, | en |
| dc.relation.page | 84 | |
| dc.identifier.doi | 10.6342/NTU202200383 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2022-02-11 | |
| 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 |
| dc.date.embargo-lift | 2027-02-10 | - |
| 顯示於系所單位： | 森林環境暨資源學系 | |
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