Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/63288
標題: Molecular Characterization and Practical Applications of a Novel Prokaryotic Promoter of Geminiviruses
雙生病毒新型原核性啟動子之分子特性分析及其應用
作者: 胡仲祺
關鍵字: 植物保護類;基礎研究;Geminiviruses;雙生病毒;豆類金黃嵌紋病毒屬;霍香薊黃脈病毒;啟動子;Begomovirus;Ageratum yellow vein virus;Promoter;Prokaryotic promoter
摘要: 
The family Geminiviridae consists of leafhopper-, treehopper- or whitefly-transmitted plantgeminiviruses that are among the most severe plant viruses in the tropics and subtropics.Packaged within geminate particles, the genomes of geminiviruses are single-strandedcircular DNAs that are replicated by a rolling circle mechanism, characteristic to prokaryotes,although their hosts are eukaryotes. Previous researches have indicated that geminivirusescan generate various DNA replication forms in bacteria, suggesting that geminivirusespossess the potential to regulate the expression of certain viral genes in prokaryotic systems.Using Ageratum Yellow Vein Virus (AYVV) as the model system, a novel prokaryoticpromoter, provisionally designated AV3, was identified at nt 615-889 of AYVV genome byour group. The aim of this proposal is to study the molecular characteristics of the AV3promoter and to apply the knowledge in the development of a novel prokaryotic geneexpression system and viral disease management. For the first year, the core region of AV3promoter and the corresponding Pribnow Box (-10, -35 region) will be fine-mapped. For thesecond year, the regulatory mechanism of the AV3 promoter and its possible role(s) in thepathogenicity of geminiviruses will be studied. For the third year, applied research will beperformed to develop the prokaryotic expression system of foreign proteins and to study thepossibility of using AV3 promoter as a target for viral disease management. Through thethree-year study, it is expected that the results obtained will shed lights on the molecularmechanisms of gene regulation in geminiviruses, and the knowledge will be further appliedin the practical researches such as the development of viral vectors and effective viraldisease management strategies.

雙生病毒是熱帶及亞熱帶地區最主要的植物病毒之ㄧ,其病毒顆粒為雙球型,內含一條或兩條的環狀單股DNA。雙生病毒感染的寄主皆為真核生物,也利用植物細胞中的DNA 複製酵素進行病毒DNA 合成。然而,雙生病毒在植物細胞中的複製形式卻是以原核生物特有的滾環式複製(rolling circle replication)方式進行。而先前研究指出雙生病毒亦可在原核系統進行複製。因此,原本屬於真核生物的雙生病毒基因體中亦應具有在原核系統中亦能表現的調控機制。而這種橫跨真核與原核生物的基因表現機制,即為本計畫欲深入探討的目標。本研究室先前以霍香薊黃脈病毒(Ageratum Yellow Vein Virus, AYVV)作為研究材料,發現了先前所未知的新型原核性啟動子,位於AYVV 基因體中核苷酸位置nt 615-889 的區段,並暫時命名為AV3啟動子。此啟動子可在大腸桿菌(Escherichia coli)中以構成性的方式大量表現報導基因。本計畫即擬以此AV3 啟動子為材料,以三年的時間進行此雙生病毒原核型啟動子的分子特性等基礎分析,及利用於生產外源蛋白與病毒病害防治等產業應用層面。第一年預計進行AV3 啟動子核心區域與轉錄因子辨識位置之分子定位及分子特性分析,並進行在真核生物中啟動子活性的分析。第二年預計進行此AV3 啟動子之調控機制之研究,探討其是否可受到病毒本身蛋白或宿主因子的調控,並將以感染性選殖株探討此AV3 啟動子在雙生病毒致病機制中的可能功能。第三年擬進行應用性研究,將研發利用AV3 啟動子核心區域表現外源蛋白之原核系統載體,並探討將AV3 啟動子作為病毒病害防制目標的可能性。預計本研究所獲得的成果將有助於瞭解雙生病毒的基因調控及其致病機制,並可進一步應用於原核型外源蛋白表現載體的開發與雙生病毒病害防治之實用層面。
URI: http://hdl.handle.net/11455/63288
其他識別: NSC97-2313-B005-038-MY3
Appears in Collections:生物科技學研究所

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