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Rearrangements of large-insert T-DNAs in transgenic rice
Authors:Akiko?Nakano  Email author" target="_blank">Go?SuzukiEmail author  Maki?Yamamoto  Kym?Turnbull  Sadequr?Rahman  Yasuhiko?Mukai
Institution:(1) Laboratory of Plant Molecular Genetics, Division of Natural Science, Osaka Kyoiku University, 4-698-1 Asahigaoka, Kashiwara Osaka, 582-8582, Japan;(2) Faculty of Health Sciences for Welfare, Kansai University of Welfare Sciences, Osaka, 582-0026, Japan;(3) CSIRO Plant Industry, P.O. Box 1600, ACT, 2601, Australia
Abstract:Introduction of large-DNA fragments into cereals by Agrobacterium-mediated transformation is a useful technique for map-based cloning and molecular breeding. However, little is known about the organization and stability of large fragments of foreign DNA introduced into plant genomes. In this study, we produced transgenic rice plants by Agrobacterium-mediated transformation with a large-insert T-DNA containing a 92-kb region of the wheat genome. The structures of the T-DNA in four independent transgenic lines were visualized by fluorescence in situ hybridization on extended DNA fibers (fiber FISH). By using this cytogenetic technique, we showed that rearrangements of the large-insert T-DNA, involving duplication, deletion and insertion, had occurred in all four lines. Deletion of long stretches of the large-insert DNA was also observed in Agrobacterium.
Keywords:Agrobacterium-mediated transformation  Fluorescence in situ hybridization on extended DNA fibers (fiber FISH)  Oryza sativa  DNA rearrangement  Large T-DNA inserts
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