2007-04-112024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/684295摘要：蘭花為台灣重要之經濟作物，具備各種消費者喜好花形、花色之重要性狀，然而 許多負責蘭花重要性狀之基因並未被充分了解及利用。本研究擬以轉位子建立蘭 花之有用基因釣取（gene tagging/trapping）系統，有助於篩選出蘭花之有用基因 及其啟動子，將來並可應用於產業快速建立蘭花之新品種。申請人先前曾建構「 可誘導轉位子」成功應用於水稻、蕃茄及菸草等作物。然而，最初之「可誘導轉 位子」僅適用於基因篩選，需大量製造同源突變體。先前，申請人完成一新構築 ：將一無啟動子之報導基因（promoter-less reporter gene）建構於先前之「可誘導 轉位子」中，已獲18株抗篩選標記（hygromycin）之轉基因蝴蝶蘭品系，該構築 轉入作物則不需製造同源突變體即可開始篩選有用基因。本研究已水楊酸誘導轉 基因蝴蝶蘭，更進一步證明該「可誘導轉位子」可在蝴蝶蘭組織中轉位，且以南 方吸漬法證明轉位子插入染色體特定位置。據此，本年度將研究轉位樣式，配合 最新之activation tagging構築開發蘭花新品種。<br> Abstract: Orchid is an important industry in Taiwan. In order to sustain and expand the market, genetic engineering has been recognized as an approach to overcome the limitations of improvement through conventional breeding. Through genetic engineering, genes for shelf life, flower color and architecture can be directly manipulated to develop new varieties that are tailor made to customers' preferences. However, many Phalaenopsis genes remained to be cloned and engineered. In this study, we will use the inducible transposon to clone the useful genes of Phalaenopsis and furthermore study their function. Transposon has been demonstrated to be a powerful tool for cloning plant genes. Thus, we have constructed an inducible transposon (INAc) and demonstrated that it is functional in rice, tomato and tobacco plants. However, by using the INAc, most plant genes could be cloned only by creating homozygous mutants. This feature limits the application of INAc for cloning genes from many important plants, e.g. Phalaenopsis. Firstly, we have constructed a new inducible transposon (termed as INAcGFP) containing a promoter-less reporter gene (GFP). This allows us to clone the useful gene from T0 generation of transgenic Phalaenopsis plant. Furthermore, the promoter-less GFP gene was replaced by 5x 35S enhancer, which located by 3' end of INAc. In this report, the new construct has been transformed into Phalaenopsis plant. The transposition patterns of transformed orchid plants containing INAcGFP were analyzed by PCR and DNA blot techniques. In this project, we will analyze the transposition pattern in Phalaenopsis to improve the Activation tagging mutants creation.可誘導轉位子轉基因植物轉基因蝴蝶蘭基因功能強化inducible transposontransgenic planttransgenic Phalaenopsisactivation tagging