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Construction of an inducible transposon,INAc, to develop a gene tagging system in higher plants
Authors:Charng  Yuh-Chyang  Pfitzner  Artur JP  Pfitzner  Ursula M  Charng-Chang  Kai-Fay  Chen  Chui-mei  Tu  Jenn  Kuo  Tsong-Teh
Institution:(1) Institute of Botany, Taiwan;(2) Present address: Department of Agronomy, National Taiwan University, Taipei, Taiwan;(3) Botanisches Institut der Ludwig-Maximilians-Universität, München, Germany;(4) Present address: Institut für Genetik, Universität Hohenheim, Stuttgart, Germany;(5) Institute of Molecular Biology, Academia Sinica, Taipei, 11529, Taiwan;(6) Graduate Institute of Botany, Taiwan
Abstract:An inducible transposable element, termed INAc (inducible Activator), was constructed for development of a gene tagging system in higher plants. The advantage of such an inducible element is that, unlike the native transposon, its excision can be induced at any time during plant development and the resulting mutants are stable after removal of the inducer. A fusion of the SA inducible promoter (PR-1a) with the Ac transposase gene was inserted together with a hygromycin resistance gene between ca. 400 bp sequences from each end of the maize Ac element, yielding INAc. The INAc element was introduced into tobacco and tomato plants. A high frequency of spontaneous transposition was apparent in primary transformed tomato calli but not in tobacco calli. Treatment of tobacco plants with salicylic acid induced transposition of INAc in both somatic and germinal tissue, with germinal transposition events being revealed by characterization of the progeny of transformed plants whose flowers were exposed to SA. The INAc element thus exhibits potential for development of an inducible transposon system suitable for gene isolation in heterologous plant species.
Keywords:Ac transposase  LUC reporter gene  PR-1a promoter  salicylic acid  transposon tagging
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