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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 蔡皇龍 | |
dc.contributor.author | Tzu-Hsien Yang | en |
dc.contributor.author | 楊子嫻 | zh_TW |
dc.date.accessioned | 2021-06-17T07:06:39Z | - |
dc.date.available | 2020-08-18 | |
dc.date.copyright | 2019-08-18 | |
dc.date.issued | 2019 | |
dc.date.submitted | 2019-07-24 | |
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AGROBEST: an efficient Agrobacterium-mediated transient expression method for versatile gene function analyses in Arabidopsis seedlings. Plant methods 10, 19. Yun, H., Hyun, Y., Kang, M.-J., Noh, Y.-S., Noh, B., and Choi, Y. (2011). Identification of regulators required for the reactivation of FLOWERING LOCUS C during Arabidopsis reproduction. Planta 234, 1237-1250 Zuo, J., Niu, Q.W., and Chua, N.H. (2000). An estrogen receptor‐based transactivator XVE mediates highly inducible gene expression in transgenic plants. The Plant Journal 24, 265-273. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72803 | - |
dc.description.abstract | 阿拉伯芥(Arabidopsis thaliana)藉由外在環境與內在基因調控開花時機,在開花途徑的調控機制中,FLOWERING LOCUS C (FLC) 為主要抑制開花的基因,植物在內外條件適合時,會透過抑制 FLC 的表現來啟動開花程序,進入繁殖階段。植物抑制FLC基因表現的途徑主要為外在長期低溫造成的春化作用(vernalization)及內在的自主控制 (autonomous control)。春化作用指植物歷經長期的低溫時,FLC基因座會受多梳抑制蛋白複合體(Polycomb Repressive Complexes)引發不可逆的組蛋白甲基化(histone methylation)造成FLC靜默(silencing),開花基因不再受抑制進而啟動開花。自主開花途徑(autonomous pathway)中,FLC基因座則經由未編碼核糖核酸(non-coding RNA)所參與的機制引發組蛋白甲基化的靜默調控。上述FLC的抑制機制皆是作用於基因座上負向調控,目前仍未發現有任何開花途徑經由FLC啟動子區域完成該基因的抑制。前人研究中發現,阿拉伯芥bzip16為晚開花突變株,造成晚開花的主因則為突變株中的FLC表現量顯著地提高,另一方面已知bZIP16能結合於FLC啟動子區域,推論bZIP16可能為FLC啟動子的抑制者。為進一步探討bZIP16在開花途徑中的角色,本論文建立bZIP16過量表現的誘導系統以及FLC啟動子的報導構築,後續將可利用農桿菌感染暫時性導入阿拉伯芥小苗中表現,觀察bZIP16對於FLC啟動子的調控角色。另外,已知FLC啟動子區域中具有bZIP16結合的保留性序列G-box (CACGTG),本論文亦建立能反映此G-box於FLC啟動子中之調控角色的報導構築,有利後續研究工作。 | zh_TW |
dc.description.abstract | The flowering time of Arabidopsis thaliana is regulated by the crosstalk of environmental and endogenous factors. FLOWERING LOCUS C (FLC), is the main repressor of flowering in Arabidopsis. Once the plant meets suitable conditions to propagate, FLC would be repressed and the flowering genes are activated. The vernalization and autonomous control, are two negative pathways inhibit the expression of FLC. Under the vernalization, the prolonged duration of low temperature silences FLC expression via Polycomb Repression Complexes (PRCs)-mediated irreversible histone methylation. In autonomous pathway, the FLC is silenced by histone methylation mediated by non-coding RNAs. The above repressive mechanisms action on the various regions of FLC locus, currently, if the FLC could be repressed via the promoter region remains unknown. It has been shown that the bzip16 mutant is late-flowered due to the enhancement of FLC expression. Also, bZIP16 associates with the FLC promoter in vivo, therefore, bZIP16 could be a repressor acting on the FLC promoter. To study the regulatory role of bZIP16 in flowering time control, this study generates an inducible system for bZIP16 overexpression and the reporter construct for FLC promoter. Furthermore, a bZIP16-binding motif, G-box (CACGTG), is found in the region that bZIP16 associated with the FLC promoter, we also generated a reporter construct to investigate the role of G-box in the promoter. All the effector and reporter constructs are established in binary vectors that will be further applied to transient assays and generate transgenic plants. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T07:06:39Z (GMT). No. of bitstreams: 1 ntu-108-R06b43032-1.pdf: 2959966 bytes, checksum: 966583ad1e6285536d0e116ec14fca7b (MD5) Previous issue date: 2019 | en |
dc.description.tableofcontents | 口試委員審定書…………………………………………………………I
致謝…………………………………………………………………………………II 摘要…………………………………………………………………………………II ABSTRACT……………………………………………………………………IV 第一章 前言 1 1.1 抑制FLC的機制 1 1.2 植物轉錄因子bZIP16可能參與FLC調控 3 1.3 研究動機與目的 4 第二章 材料與方法 5 2.1材料 5 2.1.1 植物材料 5 2.1.2基因表現質體構築 5 2.2實驗方法 5 2.2.1阿拉伯芥幼苗暫時性表現方法AGROBEST 5 2.2.2阿拉伯芥幼苗蛋白質製備 8 2.2.3西方墨點法(Western blot) 9 2.2.4 聚合連鎖反應(Polymerase chain reaction, PCR)11 2.2.5菌落聚合連鎖反應(colony PCR) 12 2.2.6 DNA連接(DNA ligation) 13 2.2.7質體DNA萃取 13 2.2.8大腸桿菌勝任細胞之轉化作用 14 2.2.9農桿菌質體轉化 14 2.2.10 DNA限制酶切位檢查 15 第三章 實驗結果 16 3.1 bZIP16誘導載體構築 16 3.2 FLC啟動子報導載體構築 16 3.3 AGROBEST( Agrobacterium-mediated enhanced seedling transformation)條件測試 17 3.4 bZIP16-GFP 過量表現可於植物細胞中被誘導 18 3.5 測試bZIP16對FLC啟動子之作用 18 3.6 bZIP16結合序列G-box報導載體構築 19 3.7 測試bZIP16結合序列G-box對FLC啟動子之作用 19 第四章 討論 21 4.1 農桿菌感染濃度對bZIP16誘導表現無明顯影響 21 4.2 bZIP16-GFP分佈情形符合核蛋白要件 21 4.3 bZIP16對於FLC啟動子調控尚未能有定論 22 4.4 bZIP16對於FLC啟動子內G-box序列之調控 23 4.5 後續研究 24 第四章 圖表 25 參考文獻 41 | |
dc.language.iso | zh-TW | |
dc.title | 轉錄因子bZIP16對開花基因FLOWERING LOCUS C啟動子之調控角色 | zh_TW |
dc.title | The role of bZIP16 in of FLOWERING LOCUS C regulation | en |
dc.type | Thesis | |
dc.date.schoolyear | 107-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳賢明,林盈仲 | |
dc.subject.keyword | 春化作用,自主性開花途徑,FLOWERING LOCUS C (FLC),bZIP16,AGROBEST, | zh_TW |
dc.subject.keyword | vernalization,autonomous flowering,FLOWERING LOCUS C (FLC),bZIP16,AGROBEST, | en |
dc.relation.page | 44 | |
dc.identifier.doi | 10.6342/NTU201901856 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2019-07-25 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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