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A one-time inducible transposon for creating knockout mutants
Authors:Kuan-Te Li  Ya-Lin Lin  Ji-Ying Huang  Wen-Ya Li  Yuh-Chyang Charng
Institution:(1) Department of Agronomy, National Taiwan University, No.1 Sec.4 Roosevelt Rd., Taipei, Taiwan, Republic of China
Abstract:The maize transposon Ac can move to a new location within the genome to create knockout mutants in transgenic plants. In rice, Ac transposon is very active but sometimes undergoes further transposition and leaves an empty mutated gene. Therefore, we developed a one-time transposon system by locating one end of the transposon in the intron of the Ac transposase gene, which is under the control of the inducible promoter (PR-1a). Treatment with salicylic acid induced transposition of this transposon, COYA, leading to transposase gene breakage in exons. The progeny plants inheriting the transposition events become stable knockout mutants, because no functional transposase could be yielded. The behavior of COYA was analyzed in single-copy transgenic rice plants. We determined the expression of the modified transposase gene and its ability to trigger transposition events in transgenic rice plants. The COYA element thus exhibits potential for development of an inducible transposon system suitable for gene isolation in heterologous plant species. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Keywords:Ac transposase  Inducible transposon  Knockout mutant  Transposon tagging
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