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RESEARCH PAPERS ON SYSTEMS BIOLOGY/GENOMICS/BIOINFORMATICS

A Resource of Mapped Dissociation Launch Pads for Targeted Insertional Mutagenesis in the Arabidopsis Genome¹

Department of Cell and Developmental Biology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom (P.R.M., L.C., A.M., C.D.); and Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 (P.S.S., R.M.)

We describe a new resource for targeted insertional mutagenesis in Arabidopsis using a maize (Zea mays) Activator/Dissociation (Ds) two-element system. The two components of the system, T-DNA vectors carrying a Ds launch pad and a stable Activator transposase source, were designed to simplify selection of transposition events and maximize their usefulness. Because Ds elements preferentially transpose to nearby genomic sites, they can be used in targeted mutagenesis of linked genes. To efficiently target all genes throughout the genome, we generated a large population of transgenic Arabidopsis plants containing the Ds launch pad construct, identified lines containing single Ds launch pad inserts, and mapped the positions of Ds launch pads in 89 lines. The integration sites of the Ds launch pads were relatively evenly distributed on all five chromosomes, except for a region of chromosomes 2 and 4 and the centromeric regions. This resource therefore provides access to the majority of the Arabidopsis genome for targeted tagging.

¹ This work was supported by the Biotechnology and Biological Sciences Research Council (grant no. PAG04435) and by the European Commission (grant no. BIO4 CT98 5004).

² Present address: Max-Planck-Institute for Plant Breeding Research, Department of Plant-Microbe Interactions, Carl von Linne Weg 10, D-50829 Cologne, Germany.

³ Present address: Istituto di Genetica Vegetale, Universita Cattolica S.C., 29100 Piacenza, Italy.

⁴ Present address: Department of Botany and Plant Science, Center for Plant Cell Biology, University of California, Riverside, CA 92521.

^* Corresponding author; e-mail muskett{at}mpiz-koeln.mpg.de; fax 49–221–5062–353.

Received October 28, 2002; returned for revision December 21, 2002; accepted January 26, 2003.

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