Expression analysis of gene trap lines and mapping of donor loci for Dissociation transposition in Arabidopsis
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In order to establish a system for characterizing gene function utilizing information obtained from genome sequences, we generated T-DNA insertion lines using a newly constructed binary vector. The vector carries a <i>uidA</i> [β-glucuronidase (<i>GUS</i>)] reporter gene which allows the promoter activity of the inserted genes to be monitored, a transposable element <i>Dissociation</i> (<i>Ds</i>) for targeted insertional mutagenesis, and the <i>cis</i> sequences required for <i>Agrobacterium</i>-mediated transformation. Approximately 8% of the 20,000 lines tested for GUS activity exhibited positive staining. Staining was detected in various organs including roots, leaves, stems, flowers, and siliques. These lines are therefore useful resources for analyzing tissue or organ specific gene expression. We have included the GUS expression patterns on our web site (http://www.kazusa.or.jp/ja/plant/GUS/). To establish a system for targeted insertional mutagenesis, integration sites of the T-DNAs in 140 lines with a single T-DNA insertion were mapped on the genome. The T-DNA contains the <i>Ds</i> element; therefore these 140 lines could be used as donor loci for <i>Ds</i> transposition. The integration sites were almost evenly distributed on all five chromosomes except for the nuclear organizer regions of chromosomes 2 and 4 and the centromeric regions. The <i>Ds</i> element in one line was transposed in combination with an <i>Activator</i> (<i>Ac</i>) element. In about half of the transposed lines, the <i>Ds</i> elements were reinserted within about 1 M bp from the donor locus. These results indicate that the donor loci for <i>Ds</i> transposition that were mapped in this study are a valuable resource for targeted mutagenesis throughout the <i>Arabidopsis</i> genome.
- Plant tissue culture letters
Plant tissue culture letters 24(5), 467-479, 2007-12-01
Japanese Society for Plant Cell and Molecular Biology