| 研究生: |
洪彧丞 Hung, Yu-Cheng |
|---|---|
| 論文名稱: |
利用染色質免疫沉澱測序(ChIP-seq)實驗數據建立植物轉錄因子結合位置資料庫 Construction of a database for transcription factor binding sites identified by plant chromatin immunoprecipitation sequencing (ChIP-seq) experiments |
| 指導教授: |
張文綺
Chang, Wen-Chi |
| 學位類別: |
碩士 Master |
| 系所名稱: |
生物科學與科技學院 - 熱帶植物科學研究所 Institute of Tropical Plant Sciences |
| 論文出版年: | 2018 |
| 畢業學年度: | 106 |
| 語文別: | 中文 |
| 論文頁數: | 74 |
| 中文關鍵詞: | 轉錄因子 、轉錄因子結合位置 、基因調控 、染色質免疫沉澱測序 、PCBase |
| 外文關鍵詞: | TFs, TFBSs, gene regulatory mechanism, ChIP-seq, PCBase |
| 相關次數: | 點閱:113 下載:0 |
| 分享至: |
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植物在不同的生長發育階段或遭遇逆境時,都需要調節相關基因的表現量來幫助自身適應不同的環境。其中最重要的調控方式就是利用轉錄因子(transcription factors)結合到啟動子(promoter)上的近端調控因子(cis-elements)來控制下游基因的表現。因此,若想了解其中複雜的調控關係,研究轉錄因子和基因組(genome)上的結合位置 (transcription factor binding sites,簡稱TFBSs)顯然是一個相當重要的工作。近年來各種高通量技術蓬勃發展,其中染色質免疫沉澱測序(chromatin immunoprecipitation sequencing,簡稱ChIP-seq)被廣泛應用於分析整個基因組上可能的轉錄因子結合位置或尋找會被某個轉錄因子所調控的基因群。所以,若能將這些實驗資料進行全面且條理性的分析統整,勢必更能了解轉錄因子與基因之間的調控關係。到目前為止,只有3個公開資料庫有提供植物ChIP-seq實驗資料,分別為PCSD、ChIPBase v2.0和Expresso。但ChIPBase v2.0和Expresso分別只收錄來自阿拉伯芥(Arabidopsis thaliana)的26和20種轉錄因子。此外,PCSD也只提供關於阿拉伯芥、水稻(Oryza sativa)和玉米(Zea mays)三種植物的轉錄因子和其調控基因的基本資訊,使用者無法直接分析任意啟動子序列上是否存在實驗驗證的TFBSs。基於以上原因,本研究主要的目的為建立一個整合多種植物物種之ChIP-seq資料庫。我們由Gene Expression Omnibus (GEO)和Sequence Read Archive (SRA)中蒐集了阿拉伯芥、水稻、玉米、大豆(Glycine max)、番茄(Solanum lycopersicum)、棉花(Gossypium hirsutum)和深山南芥(Arabidopsis lyrata)共7種植物之ChIP-seq實驗資料,涵蓋662個samples和99種蛋白質(包含轉錄因子、組織蛋白和其他DNA結合蛋白)。以上所有資料經由系統性的流程處理,並由AQUAS及MACS2分析,最終獲得4,574,337筆蛋白質結合位置以及1,233,999筆蛋白質與基因之結合關係資訊。最後建立一個名為Plant ChIP-seq Database(簡稱PCBase)的資料庫以方便植物學家快速了解基因組DNA上多種蛋白質的結合位置。PCBase提供基因搜尋(Gene Search),蛋白質搜尋(Protein Search),啟動子分析(Promoter Analysis),基因組瀏覽(Genome Browser)及資料下載(Download)等五大功能來全面分析植物染色質狀態。使用者除了能輕易獲得轉錄因子和其轉錄因子結合位置座標及調控基因外,所有由ChIP-seq實驗資料得來的轉錄因子結合位置權重(TF binding position weight matrices)皆可用於分析任一段啟動子序列。我們希望PCBase可以幫助植物領域研究人員在研究植物的轉錄調控機制時更加便利。目前PCBase可由以下網址免費取得:http://pcbase.itps.ncku.edu.tw/
Transcription factors (TFs) can bind to cis-elements on promoters and regulate gene expression during plant development and stresses responsiveness. Therefore, investigation of TFs and their binding sites (TFBSs) is important to understand the gene regulatory mechanisms. Recently, a high-throughput technology, chromatin immunoprecipitation sequencing (ChIP-seq), was applied to rapidly explore the binding sequences on a genome and regulatory genes of a TF in many studies. Consequently, the collection and analysis of ChIP-seq experimental data will be useful to further identify the relationships among TFs, TFBSs, and TF-targeted genes. Until now, only three databases provide ChIP-seq data from plants, those are PCSD (including Arabidopsis thaliana, Oryza sativa (rice), Zea mays (maize)), ChIPBase v2.0 and Expresso (including Arabidopsis thaliana), respectively. However, only 26 and 20 TFs can be accessed from ChIPBase v2.0 and Expresso, individually. Additionally, PCSD only provides the information of TFs and their target-genes; the experimental TF binding motifs can’t be applied in promoter analysis. Based on the above, the aim of this study is to construct a comprehensive plant ChIP-seq database. We collected ChIP-seq experimental data from seven plants including Arabidopsis thaliana, Oryza sativa (rice), Zea mays (maize), Glycine max (soybean), Solanum lycopersicum (tomato), Gossypium hirsutum (cotton) and Arabidopsis lyrata. The number of samples obtained from Gene Expression Omnibus (GEO) and Sequence Read Archive (SRA) are 662, which contain 99 proteins (TF/Histone/DNA binding protein). AQUAS and MACS2 programs systematically processed all of the raw data to identify the protein binding sites. Finally, we constructed a database named Plant ChIP-seq Database (PCBase) to comprehensively analyze plant chromatin status. There are five main functions in PCBase, such as Gene Search, Protein Search, Promoter Analysis, Genome Browser, and Download. The target genes and TFBS coordinates of a TF can be effectively retrieved. All experimental TF binding matrices can be utilized in any input promoter sequence. We hope this database can help scientists to study the transcriptional regulatory mechanisms in plants. This database is freely available at: http://pcbase.itps.ncku.edu.tw/
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校內:2023-08-27公開