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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林詩舜 | zh_TW |
| dc.contributor.advisor | Shih-Shun Lin | en |
| dc.contributor.author | 阮香江 | zh_TW |
| dc.contributor.author | Huong Giang Nguyen | en |
| dc.date.accessioned | 2026-08-11T16:20:33Z | - |
| dc.date.available | 2026-08-12 | - |
| dc.date.copyright | 2026-08-11 | - |
| dc.date.issued | 2026 | - |
| dc.date.submitted | 2026-08-03 00:00:00 | - |
| dc.identifier.citation | References
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/103389 | - |
| dc.description.abstract | 目前針對 HC-Pro 介導 RNA 沉默抑制之分子機制研究，仍受限於適當的實驗工具與分析方法。本實驗室先前研究已證實，來自蕪菁嵌紋病毒（turnip mosaic virus, TuMV）的 P1/HC-Pro 蛋白（P1/HC-ProTu）可有效抑制 HEN1 的活性，導致未甲基化 miRNA（unmethylated miRNAs, unMet-miRNAs）累積，並進一步誘導 HESO1 介導的 AGO1 自噬性降解。為建立後續機制研究所需的平台，本研究建立了一套完整的研究系統，包括可作為蛋白質定量內部對照之可靠 ACTIN 抗體、可精確控制 HC-ProTu 時間性表達的誘導型轉基因植物（XVE–P1/HC-ProTu/hen1-8/heso1-1 plant 與 XVE–P1/HC-ProTu/Col-0 plant），以及結合 ContigViews 的轉錄體基因調控網路分析流程，用於解析 RNA 沉默途徑受干擾後所引起的基因表現變化。整體而言，本研究建立了一套整合性的研究平台，可作為未來探討病原如何操控小 RNA 調控途徑，以及 RNA 沉默失調如何重塑植物基因調控網路的重要基礎。 | zh_TW |
| dc.description.abstract | Mechanistic studies of HC-Pro-mediated RNA silencing disruption remain limited by the availability of suitable experimental tools and analytical approaches. Our previous study demonstrated that the P1/HC-Pro protein from turnip mosaic virus (P1/HC-ProTu) is a potent inhibitor of HEN1, leading to the accumulation of unmethylated miRNAs (unMet-miRNAs) and triggering HESO1-mediated autophagic degradation of AGO1. To provide a foundation for future investigations, this study established a research platform consisting of a reliable ACTIN antibody for protein normalization, inducible HC-ProTu transgenic lines including XVE–P1/HC-ProTu/hen1-8-heso1-1, XVE–P1/HC-ProTu/Col-0 plants that allow precise temporal control of HC-ProTu expression, and a transcriptome-based gene network analysis workflow using ContigViews to investigate changes in gene expression associated with perturbation of the RNA silencing pathway. Together, these resources establish an integrated study system that provides a foundation for future studies aimed at understanding how pathogens manipulate small RNA pathways and how RNA silencing disruption reshapes plant gene regulatory networks. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2026-08-11T16:20:33Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2026-08-11T16:20:33Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | Contents
Acknowledgements i 摘要 ii Abstract iii Contents iv List of tables vi List of figures vii Introduction 1 Materials and Methods 5 Plant material and growth conditions 5 Generation for inducible P1/HC-ProTu plants 5 Recombinant protein purification 6 Western blot 7 Beta-elimination and northern blot for Met- and unMet-miRNA detection 8 Comparative network analysis by ContigViews 9 Results 11 A homemade ACTIN antibody can detect endogenous ACTIN 11 Inducible transgenic plants show a mimic phenotype of wildtype and mutants 13 Induced P1/HC-ProTu can lead to AGO1 degradation 14 Induced P1/HC-ProTu can interfere with miRNA methylation 15 Comparative gene-to-gene network and transcriptome analysis for data mining 16 Discussion 19 Homemade ACT7-derived antibody can stable detect endogenous actin as a reliable internal control 19 An inducible P1/HC-ProTu system establishes P1/HC-ProTu as the direct cause of AGO1 destabilization and impaired miRNA methylation 19 HC-Pro-mediated RNA silencing disruption may reshape growth–defense regulatory networks 21 Application of the developed experimental tools for further investigations. 22 Conclusion 24 References 25 Tables 34 Figures 42 Supplementary 54 | - |
| dc.language.iso | en | - |
| dc.subject | RNA 沉默 | - |
| dc.subject | HC-ProTu | - |
| dc.subject | miRNA 甲基化 | - |
| dc.subject | ACTIN 抗體 | - |
| dc.subject | 基因調控網路分析 | - |
| dc.subject | RNA silencing | - |
| dc.subject | HC-ProTu | - |
| dc.subject | miRNA methylation | - |
| dc.subject | antibody | - |
| dc.subject | gene network analysis | - |
| dc.title | 阿拉伯芥中 HC-Pro 介導 RNA 靜默抑制之研究工具 | zh_TW |
| dc.title | Development of a toolkit for the analysis of HC-Pro-mediated RNA silencing suppression in Arabidopsis thaliana | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 114-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 陳荷明;張碧芳; 呂冠儒; 唐涵 | zh_TW |
| dc.contributor.oralexamcommittee | Ho-Ming Chen;Pi-Fang Chang;Kuan-Ju Lu;Han Tang | en |
| dc.subject.keyword | RNA 沉默; HC-ProTu; miRNA 甲基化; ACTIN 抗體; 基因調控網路分析 | zh_TW |
| dc.subject.keyword | RNA silencing; HC-ProTu; miRNA methylation; antibody; gene network analysis | en |
| dc.relation.page | 55 | - |
| dc.identifier.doi | 10.6342/NTU202602608 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2026-08-04 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 生物科技研究所 | - |
| dc.date.embargo-lift | 2026-08-12 | - |
| 顯示於系所單位： | 生物科技研究所 | |
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