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A set of modular plant transformation vectors allowing flexible insertion of up to six expression units

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Abstract

We have constructed a binary vector for Agrobacterium-mediated plant transformation, which has a multiple cloning site consisting of 13 hexanucleotide restriction sites, 6 octanucleotide restriction sites and 5 homing endonuclease sites. The homing endonuclease sites have the advantages to be extremely rare in natural sequences and to allow unidirectional cloning. We have also constructed a set of auxiliary vectors allowing the assembly of expression cassettes flanked by homing endonuclease sites. The expression cassettes assembled in these auxiliary vectors can be transferred into the binary vector with virtually no risk of cutting the vector within previously introduced sequences. This vector set is ideally suited for the construction of plant transformation vectors containing multiple expression cassettes and/or other elements such as matrix attachment regions. With this modular vector system, six different expression units were constructed in as many auxiliary vectors and assembled together in one plant transformation vector. The transgenic nature of Arabidopsis thaliana plants, transformed with this plant transformation vector, was assessed and the expression of each of the six genes was demonstrated.

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Author notes

  1. Willem F. Broekaert

    Present address: Crop-Design N.V., Technologiepark 3, 9052, Gent, Belgium

Authors and Affiliations

  1. Centrum voor Microbiële en Plantengenetica (formerly F.A. Janssens Laboratorium voor Genetica), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, 3001, Heverlee-Leuven, Belgium

    Inge J.W.M. Goderis, Miguel F.C. De Bolle, Isabelle E.J.A. François, Piet F.J. Wouters, Willem F. Broekaert & Bruno P.A. Cammue

  2. Flanders Interuniversity Institute for Biotechnology – PRJ3, Kasteelpark Arenberg 20, 3001, Heverlee-Leuven, Belgium

    Inge J.W.M. Goderis, Miguel F.C. De Bolle, Piet F.J. Wouters & Bruno P.A. Cammue

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  1. Inge J.W.M. Goderis
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Goderis, I.J., De Bolle, M.F., François, I.E. et al. A set of modular plant transformation vectors allowing flexible insertion of up to six expression units. Plant Mol Biol 50, 17–27 (2002). https://doi.org/10.1023/A:1016052416053

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