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

Author:

Muskett Paul R.1,Clissold Leah1,Marocco Adriano1,Springer Patricia S.1,Martienssen Robert1,Dean Caroline1

Affiliation:

1. 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.)

Abstract

Abstract 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.

Publisher

Oxford University Press (OUP)

Subject

Plant Science,Genetics,Physiology

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