Characterization and mapping of Ds—GUS-T-DNA lines for targeted insertional mutagenesis |
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| Authors: | D Smith Y Yanai Y-G Liu S Ishiguro K Okada D Shibata RF Whittier NV Fedoroff |
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| Institution: | Carnegie Institution of Washington, Department of Embryology, 115 West University Parkway, Baltimore, MD 21210, USA;Mitsui Plant Biotechnology Research Institute, Sengen 2-1-6 TCI-D21 Tsukuba, Ibaraki 305, Japan;National Institute for Basic Biology, 38 Nishigonaka, Myodaijicho, Okazaki 444, Japan |
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| Abstract: | The transposition patterns of the Ds —GUS transposon T-DNA in 23 independent single-copy lines have been characterized and the map positions of 10 of them on three of the five Arabidopsis chromosomes are reported. Using overexpressed Activator ( Ac ) elements as a transposase source, it was found that the primary determinant of transposition frequency is the insertion site of the Ac -T-DNA. Neither the structure of the transposon T-DNA nor, in most cases, its insertion site have a significant effect on transposition frequency. Both the frequency and timing of transposition are influenced by the parent through which the transposon and transposase T-DNAs are transmitted. Overall, nearly 75% of plants in which excision has occurred bear a reinserted element and very short-range transpositions predominate, underlining the advantage of using mapped transposons for insertional mutagenesis. |
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