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Agrobacterium tumefaciens T-DNA integrates into multiple sites of the sunflower crown gall genome

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Summary

We analyzed the integration of a tumor inducing (Ti) plasmid into an octopine producing crown gall tumor of sunflower, line PSCG 15955. A continuous Ti plasmid segment (T-DNA) of about 19.5 kilo base pairs (kbp) is transferred and integrated into a small number of sites of the plant DNA.

The number of T-DNA integration sites in our tumor line was estimated by two different methods. First, cloned fragments of the T-DNA were hybridized to tumor DNA and the hybridization patterns were observed. The number of T-DNA integration sites in line PSCG 15955 was found to be approximately eight.

Second, a library was constructed from total DNA of tumor line PSCG 15955 by molecular cloning using the bacteriophage lambda vector Charon 4A. Recombinant phages having sequence homologies to the Ti plasmid were selected. The lower limit on the number of integration sites was three because we obtained three different right hand side genomic clones of plant/T-DNA hybrids. The initial screening of the library also revealed two left hand border clones. Hybridization of these five distinct recombinant clones to uninfected sunflower DNA shows that the cloned T-DNA segments are covalently bonded to plant DNA. The left and right hand plant boundary sequences are homologous to either unique and or repeated plant DNA segments.

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Author notes

  1. Doris Ursic

    Present address: Doris Ursic, Department of Genetics, University of Wisconsin, 53706, Madison, Wisconsin, USA

Authors and Affiliations

  1. Plant Disease Resistance Research Unit, ARS, SEA, USDA Department of Plant Pathology, University of Wisconsin, 53706, Madison, Wisconsin, USA

    Doris Ursic & John D. Kemp

  2. Plant Disease Resistance Research Unit, ARS, SEA, USDA Department of Genetics, University of Wisconsin, 53706, Madison, Wisconsin, USA

    Jerry L. Slightom

Authors
  1. Doris Ursic
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  2. Jerry L. Slightom
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  3. John D. Kemp
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Communicated by F. Kaudewitz

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Ursic, D., Slightom, J.L. & Kemp, J.D. Agrobacterium tumefaciens T-DNA integrates into multiple sites of the sunflower crown gall genome. Mol Gen Genet 190, 494–503 (1983). https://doi.org/10.1007/BF00331082

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  • DOI: https://doi.org/10.1007/BF00331082

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