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Transgenic tomato lines containing Ds elements at defined genomic positions as tools for targeted transposon tagging

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Abstract

We have introduced a genetically marked Dissociation transposable element (Ds HPT) into tomato (Lycopersicon esculentum) by Agrobacterium tumefaciens-mediated transformation. Probes for the flanking regions of the T-DNA and transposed Ds HPT elements were obtained with the inverse polymerase chain reaction (IPCR) technique and used in RFLP linkage analyses. The RFLP map location of 11 T-DNAs carrying Ds HPT was determined. The T-DNAs are distributed on 7 of the 12 tomato chromosomes. To explore the feasibility of gene tagging strategies in tomato using Ds HPT, we examined the genomic distribution of Ds HPT receptor sites relative to the location of two different, but very closely linked, T-DNA insertion sites. After crosses with plants expressing Ac transposase, the hygromycin phosphotransferase (HPT) marker on the Ds element and the excision markers β-glucuronidase (GUS) and Basta resistance (BAR) facilitated the identification of plants bearing germinally transposed Ds HPT elements. RFLP mapping of 21 transposed Ds HPT elements originating from the two different T-DNA insertions revealed distinct patterns of reintegration sites.

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Author notes

  1. Susanne Knapp

    Present address: UNILEVER Research Laboratory, Colworth House, MK44 1LQ, Sharnbrook, Bedford, UK

  2. Yvan Larondelle

    Present address: Unité de Biochimie de la Nutrition, Université Catholique de Louvain, Place Croix du Sud 2, bte 8, B-1348, Louvain-La-Neuve, Belgium

  3. Douglas Furtek

    Present address: 212 Borland Laboratory, American Cocoa Research Institute, The Pennsylvania State University, 16802-2505, University Park, PA, USA

  4. Klaus Theres

    Present address: Institut für Genetik, Carl-von-Linne-Weg 10, D-50829, Köln, Germany

Authors and Affiliations

  1. Institut für Genetik, Universität zu Köln, Weyertal 121, 50931, Köln, Germany

    Susanne Knapp, Yvan Larondelle, Mathias Roßberg, Douglas Furtek & Klaus Theres

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  1. Susanne Knapp
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  2. Yvan Larondelle
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  3. Mathias Roßberg
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  4. Douglas Furtek
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  5. Klaus Theres
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Communicated by H. Saedler

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Knapp, S., Larondelle, Y., Roßberg, M. et al. Transgenic tomato lines containing Ds elements at defined genomic positions as tools for targeted transposon tagging. Molec. Gen. Genet. 243, 666–673 (1994). https://doi.org/10.1007/BF00279576

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  • DOI: https://doi.org/10.1007/BF00279576

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