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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林盈仲 | zh_TW |
| dc.contributor.advisor | Ying-Chung Jimmy Lin | en |
| dc.contributor.author | 繆芳 | zh_TW |
| dc.contributor.author | Fang Miao | en |
| dc.date.accessioned | 2021-07-11T15:24:17Z | - |
| dc.date.available | 2024-01-23 | - |
| dc.date.copyright | 2019-01-24 | - |
| dc.date.issued | 2018 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | Aspeborg, H., Schrader, J., Coutinho, P.M. (2005) Carbohydrate-active enzymes involved in the secondary cell wall biogenesis in hybrid aspen. Plant Physiology 137: 983-997.
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(2014) Cellulose Synthesis and Its Regulation. The Arabidopsis Book 11: e0169 Li, S., Zhen, C., Xu, W., Wang, C., Cheng, Y. (2017) Simple, rapid and efficient transformation of genotype Nisqually-1: a basic tool for the first sequenced model tree. Scientific Reports 7: 2638 Lin, Y.J., et al. (2017) Reciprocal cross-regulation of VND and SND multigene TF families for wood formation in Populus trichocarpa. Proceedings of the National Academy of Sciences of the United States of Amerca. 114(45): 9722-7929 Miedes, E. et al. (2014) The role of the secondary cell wall in plant resistance to pathogens. Frontiers in plant science 5: 358 Taylor, N.G., Laurie, S., Turner, S.R. (2000) Multiple Cellulose Synthase Catalytic Subunits Are Required for Cellulose Synthesis in Arabidopsis. The plant cell.12: 2529-2539 Nilsson, J. et al. (2008) Dissecting the molecular basis of the regulation of wood formation by auxin in hybrid aspen. Plant Cell 20(4): 843-55. Novaes, E. et al. (2010) Lignin and Biomass: A Negative Correlation for Wood Formation and Lignin Content in Trees. Plant physiology 154(2): 555-561 Oh, S., Park, S., Han, K.H. (2003) Transcriptional regulation of secondary growth in Arabidopsis thaliana. Journal of Experimental Botany 54(393): 2709-2722 Pavy, N. et al. (2018) Identification of conserved core xylem gene sets: conifer cDNA microarray development, transcript profiling and computational analyses. New Phytol. 180(4): 766-86. Peter, I. S., Davidson, E.H. (2016) Chapter Thirteen - Implications of Developmental Gene Regulatory Networks Inside and Outside Developmental Biology. Current Topics in Developmental Biology. 117: 237-251. Plomion, C., Leprovost, G., Stokes, A. (2001) Wood Formation in Trees. Plant physiology 127: 1513-1523 Reece-Hoyes, J.S. et al. (2011) Enhanced yeast one-hybrid assays for high-throughput gene-centered regulatory network mapping. Nature Methods 8(12): 1059-1064 Reece-Hoyes, J.S., Marian Walhout, A.J. (2012) Yeast one-hybrid assays: A historical and technical perspective. Methods 57: 441-447 Schuetz, M., Smith, R., and Ellis, B. (2013) Xylem tissue specification, patterning, and differentiation mechanisms. Journal of Experimental Botany 64(1): 11-31. Shahmuradov, I. A., Solovyev, V. V., and Gammerman1 A. J. (2005) Plant promoter prediction with confidence estimation. Nucleic Acids Research 33(3): 1069-1076 Song, J. et al. (2006) Genetic Transformation of Populus trichocarpa Genotype Nisqually-1: A Functional Genomic Tool for Woody Plants. Plant Cell Physiology 47: 1582-1589 Sun, Y. et al. (2017) Application of the yeast one-hybrid technique to plant functional genomics studies. Biotechnology & Biotechnological Equipment 31(6): 1087-1092 Taylor, N.G. et al. (2002) Interactions among three distinct CesA proteins essential for cellulose synthesis. Proceedings of the National Academy of Sciences of the United States of Amerca.100(3): 1450-1455 Tong, A.H.Y., et al. 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(2014) Complexity of the transcriptional network controlling secondary wall biosynthesis. Plant science 229: 193-207 Zhong, R. and Ye, Z.H. (2015) Secondary cell walls: biosynthesis, patterned deposition and transcriptional regulation. Plant and Cell Physiology 56(2): 195–214. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78854 | - |
| dc.description.abstract | 基因調控網路 (gene regulatory networks, GRNs) 可以幫助人們瞭解在基因進行轉錄表達的時候調控因子 (transcription factor, TF) 和目標基因 (target gene) 之間的相互作用關係，而酵母單雜交 (yeast one-hybrid, Y1H) 系統是可以快速建立這種網路的研究方法之一。酵母單雜交的原理是分別將含有目標基因接著報導基因的質體和含有TF表達序列的質體轉殖到酵母菌中，使TF和目標DNA存在於同一個細胞裡，然後通過對這種酵母菌細胞中報導基因表達的篩選，來偵測TF和DNA是否有交互作用。在過去的研究中，酵母單雜交依照操作方式分為兩種類型：單倍體轉殖法 (haploid-transformation) 和雙倍體交配法 (diploid-mating)。
我們透過酵母單雜交系統研究木材合成相關的TF和DNA之間的結合，來初步建立木材形成相關的基因調控網路。本文以木本模式植物毛果楊為研究物種，挑選與木材形成相關的基因為研究材料。接著基於傳統酵母單雜交系統，我們引入了合成遺傳陣列 (synthetic genetic array, SGA) 技術，開發出一種適應高通量操作的新酵母單雜交系統，即減數分裂－酵母單雜交系統 (meiosis-directed Y1H system)。 通過和傳統的酵母單雜交系統進行測試對比，我們發現這種減數分裂－酵母單雜交系統，能夠進行快速簡便的高通量操作，並且得到最高的偵測結合數量，是一種可以快速高效建立基因調控網路的研究方法。 | zh_TW |
| dc.description.abstract | Gene regulatory networks (GRNs) can demonstrate the interactions between transcription factors (TFs) and their target genes, which would be detected by yeast one-hybrid (Y1H). In Y1H, two components are needed: DNA baits and TF preys, and the interactions between TF and their target DNA are measured by the downstream reporter gene activation in yeast. In the past studies, the yeast one-hybrid system was categorized into two different methods: haploid-transformation and mating diploid-mating. In our study, a high-throughput meiosis-directed Y1H system was developed by synthetic genetic array (SGA) technique. After testing meiosis-direct method, haploid-transformation and diploid-mating method together, we found that the novel method has benefits in highest discovery rate and short processing time. This meiosis-direct Y1H system can be used to quickly constructing the gene regulatory network in wood formation. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:24:17Z (GMT). No. of bitstreams: 1 ntu-107-R05b21035-1.pdf: 3836442 bytes, checksum: bf8bb02a2003a05d3f66324a5d05a6ce (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 目錄
中文摘要 I Abstract II 誌謝 III 目錄 IV 圖目錄 VII 表目錄 VIII 壹、前言 1 1.1 木材 1 1.1.1 毛果楊 1 1.1.2 木材形成 1 1.1.3 木材形成相關的基因 2 1.1.4 在其他植物中木材形成基因調控網路的研究 3 1.2 基因調控網路 4 1.2.1 轉錄因子 5 1.2.2 啟動子 5 1.3 酵母單雜交 6 1.3.1 酵母的繁殖方式 6 1.3.2 酵母單雜交原理 6 1.3.3 酵母單雜交試驗的分類 7 1.4 Synthetic genetic array (SGA) 8 1.5減數分裂酵母單雜交系統 8 貳、實驗目的 10 叁、材料與方法 11 3.1 TF和啟動子的來源 11 3.2 聚合酶鏈式反應 11 3.2.1 Pfu聚合酶鏈式反應 11 3.2.2 Taq聚合酶鏈式反應 13 3.2.3 DNA瓊脂糖凝膠電泳 14 3.2.4 切膠純化回收DNA 15 3.3 大腸桿菌轉殖和質體DNA純化 16 3.3.1 以擴增質體DNA為目的的大腸桿菌轉殖 16 3.3.2 Ligation產物之質體DNA的大腸桿菌轉殖 16 3.3.3 從大腸桿菌中純化質體DNA 17 3.4 DNA ligation反應 18 3.5 菌株來源 18 3.5.1 Y1HGold菌株 18 3.5.2 Magic Marker及相關的基因 18 3.5.3 Y1HGold-MM菌株 19 3.6 cDNA庫的建立 20 3.7 構建誘導質體 20 3.8 培養基配製 22 3.9 TF轉殖(PEG/LiAc轉殖系統) 27 3.10 TF轉殖後的單株菌落挑選 28 3.11 菌株的保存 28 3.12 構建誘導菌株 29 3.13 報導基因表達測試 29 3.14 高通量操作流程 30 3.14.1 機器和耗材 30 3.14.2 培養平盤的配製 30 3.14.3 構建96孔盤和384陣列盤 30 3.14.4 主要流程 31 3.15 偵測蛋白質-DNA結合 32 3.16 數據分析 33 3.17 構建蛋白表達質體 34 3.17.1 構建表達質體 34 3.17.2 質體的純化 34 3.17.3 TF蛋白表達 36 3.18 西方墨點法 36 肆、結果 42 4.1 質體的構建 42 4.2 轉殖酵母菌的構建和篩選 42 4.2.1 單倍體轉殖法中的酵母菌 42 4.2.2 Y1HGold-MM單倍體酵母菌 43 4.2.3 雙倍體交配法中的酵母菌 44 4.2.4 減數分裂法中的酵母菌 45 4.3 報導基因表達測試 45 4.4 構建高通量操作使用的陣列盤 46 4.5 減數分裂酵母單雜交系統和兩種傳統方法的對比 47 4.6 減數分裂法的重複性 48 4.7 構建基因調控網路 48 4.8 蛋白表達質體的構建結果 49 伍、結論 50 5.1 酵母單雜交系統的限制 50 5.2 減數分裂酵母單雜交系統的優勢 50 5.3 總結 52 陸、參考文獻 54 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 酵母單雜交 | zh_TW |
| dc.subject | 啟動子 | zh_TW |
| dc.subject | 基因調控網路 | zh_TW |
| dc.subject | 合成遺傳陣列 | zh_TW |
| dc.subject | 轉錄因子 | zh_TW |
| dc.subject | transcription factor | en |
| dc.subject | gene regulatory network | en |
| dc.subject | promoter | en |
| dc.subject | yeast one-hybrid | en |
| dc.subject | synthetic genetic array | en |
| dc.title | 使用減數分裂－酵母單雜交方法研究毛果楊中木材形成的基因調控網路 | zh_TW |
| dc.title | A meiosis-directed yeast one-hybrid system to uncover the transcriptional regulatory network in Populous trichocarpa wood formation | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 107-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 張典顯;王雅筠;周信宏 | zh_TW |
| dc.contributor.oralexamcommittee | Tien-Hsien Chang;Ya-Yun Wang;Hsin-Hung David Chou | en |
| dc.subject.keyword | 基因調控網路,轉錄因子,啟動子,酵母單雜交,合成遺傳陣列, | zh_TW |
| dc.subject.keyword | gene regulatory network,transcription factor,promoter,yeast one-hybrid,synthetic genetic array, | en |
| dc.relation.page | 85 | - |
| dc.identifier.doi | 10.6342/NTU201804193 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2019-01-16 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 生命科學系 | - |
| dc.date.embargo-lift | 2024-01-24 | - |
| 顯示於系所單位： | 生命科學系 | |
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