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Development of an efficient two-element transposon tagging system in Arabidopsis thaliana

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Summary

Modified Ac and Ds elements, in combination with dominant markers (to facilitate monitoring of excision, reinsertion and segregation of the elements) were introduced into Arabidopsis thaliana ecotype Landsberg erecta. The frequencies of somatic and germinal transactivation of the Ds elements were monitored using a streptomycin resistance assay. Transactivation was significantly higher from a stable Ac (sAc) carrying a 537 by deletion of the CpG-rich 5′ untranslated leader of the transposase mRNA than from a wild-type sAc. However, substitution of the central 1.77 kb of the transposase open reading frame (ORF) with a hygromycin resistance marker did not alter the excision frequency of a Ds element. β-Glucuronidase (GUS) or iaaH markers were linked to the transposase source to allow the identification of plants in which the transposase source had segregated away from the transposed Ds element, eliminating the possibility of somatic or germinal re-activation. Segregation of the excision marker, Ds and sAc was monitored in the progeny of plants showing germinal excision of Ds. 29% of the plants inheriting the excision marker carried a transposed Ds element.

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Author notes

  1. Christina Sjodin

    Present address: Department of Molecular Genetics, Uppsala Genetic Centre, Swedish University of Agricultural Sciences, Sweden

Authors and Affiliations

  1. AFRC, IPSR, Cambridge Laboratory, John Innes Centre, NR4 7UJ, Colney Lane, Norwich, UK

    Ian Bancroft, Anuj M. Bhatt, Christina Sjodin & Caroline Dean

  2. Sainsbury Laboratory, John Innes Centre, NR4 7UH, Colney Lane, Norwich, UK

    Steve Scofield & Jonathan D. G. Jones

Authors
  1. Ian Bancroft
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  2. Anuj M. Bhatt
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  3. Christina Sjodin
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  4. Steve Scofield
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  5. Jonathan D. G. Jones
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  6. Caroline Dean
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Communicated by D. Lonsdale

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Bancroft, I., Bhatt, A.M., Sjodin, C. et al. Development of an efficient two-element transposon tagging system in Arabidopsis thaliana . Molec. Gen. Genet. 233, 449–461 (1992). https://doi.org/10.1007/BF00265443

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  • DOI: https://doi.org/10.1007/BF00265443

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