2009-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/689075摘要：轉位子（又稱跳躍基因）已被應用於釣取一些重要之植物基因，Ac/Ds為最常用之轉位系統。先前，申請人曾將來自菸草之可誘導啟動子 PR-1a 與轉位&#37238;融合並建構於一Ds轉位子中，而成「可誘導轉位子」，轉位&#37238;之活性可受化學藥劑誘導而驅動轉位子，發現在菸草、水稻中之轉位效率約為6%，但仍出現自動轉位現象。據此，本計畫將利用來自動物（並可用於植物）之可誘導起動子驅動轉位&#37238;，建構新「可誘導轉位子」，新轉位子可誘導各種劑量轉位&#37238;，記錄其所驅動之轉位效率，應用於創造knock-out mutants。此外，申請人最近發現Ds轉位子中一個新的splice donor site，將Ds建構於抗嘉磷賽EPSPS基因之各intron中，並轉入水稻或阿拉伯芥，觀察選擇性剪接或exonization，尋找更多組splice donor/acceptor site，並可研究選擇性剪接造成Ds所插入基因產生不同mRNA之影響。<br> Abstract: Transposon has been proven to be a useful tool as an insertional mutagen for plant functional genomics studies. The aim of this project is to improve the inducible transposons based on the Ac/Ds system and alternative splicing or exonization of a gene inserted by Ds. Previously, the open reading frame coding for the transposase gene of the maize transposon Activator (Ac) was fused with the inducible promoter (PR-1a) then inserted into a Ds element to yield the inducible transposon INAc. This system is also functional in tobacco, tomato and rice plants but spontaneous transpositions were also observed. In this project, animal inducible promoters, which have been demonstrated functional in plants, would be used for constructing new inducible transposons. By inducing various amount of the transposase, the transposition efficiencies can be promoted to create knock-out mutants for functional genomics studies. Furthermore, since we have found a new splicing donor site in the Ds 5’ end, a series glyphosate-tolerant EPSPS genes inserted by Ds in each intron will be transformed into rice or Arabidopsis to study the alternative splicing or exonization effect. All of these studies will allow us understand transposon’s evolutional effects and to develop more efficient transposon systems.轉位子基因可誘導TransposonGeneinduce