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Construction and Application of Efficient Ac-Ds Transposon Tagging Vectors in Rice
Authors:Shaohong Qu  Jong-Seong Jeon  Pieter BFOuwerkerk  Maria Bellizzi  Jan Leach  Pamela Ronald  Guo-Liang Wang
Institution:( Department of Plant Pathology, Ohio State University,;Columbus OH 43210, USA;Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences,;Hangzhou 310021, China;Department of Plant Pathology, University of California,;Davis CA 95616, USA;Graduate School of Biotechnology and Plant Metabolism Research Center,;Yongin 446-701, Korea;Institute of Biology, Leiden University, Clusius Laboratory,;RA Leiden 2300, The Netherlands;Bioagricultural Sciences and Pest Management, Colorado State University,;Fort Collins, CO 80523, USA;Crop Gene Engineering Key Laboratory of Hunan Province and Pre-State Key Laboratory of Crop Germplasm Renovation and Resource Utilization, Hunan Agricultural University,;Changsha 410128, China )
Abstract:Transposons are effective mutagens alternative to T-DNA for the generation of insertional mutants in many plant species including those whose transformation is inefficient. The current strategies of transposon tagging are usually slow and labor-intensive and yield low frequency of tagged lines. We have constructed a series of transposon tagging vectors based on three approaches: (i) Ac TPase controlled by glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain (GVG) chemical-inducible expression system; (ii) deletion of Ac TPase via Cre- lox site-specific recombination that was initially triggered by Ds excision; and (iii) suppression of early transposition events in transformed rice callus through a dual-functional hygromycin resistance gene in a novel Ds element ( HPT-Ds ). We tested these vectors in transgenic rice and characterized the transposition events. Our results showed that these vectors are useful resources for functional genomics of rice and other crop plants. The vectors are freely available for the community.
Keywords:Ac-Ds transposable element  glucocorticoid binding domain/VP16 acidic activation domain/Gal4 DNA-binding domain-inducible expression  Cre-lox site-specific recombination  rice
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