Improved integration of exogenous dna delivered to plant cells

C - Chemistry – Metallurgy – 12 – N

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C12N 15/82 (2006.01) A01H 5/00 (2006.01)

Patent

CA 2230966

The present invention provides an improved method for achieving stable integration of an exogenous DNA fragment in intact form into the genome of a eukaryotic cell, particularly a plant cell. The method comprises providing the exogenous DNA together with one or more proteins which promote integration of the exogenous DNA to the eukaryotic cell targeted for transformation, wherein the proteins are provided in the form of a chimeric gene or translatable RNA capable of expression in the eukaryotic cell. The method is particularly applied to plant cells to achieve stable integration of an exogenous DNA fragment bounded by T-DNA borders in intact form using integration-promoting proteins derived from Agrobacterium. Transgenic cultures, tissues and whole organisms, particularly transgenic plants, can be generated from cells transformed according to the method of the invention.

L'invention concerne un procédé amélioré pour assurer une intégration stable d'un fragment d'ADN exogène sous une forme intacte, dans le génome d'une cellule eucaryote, en particulier d'une cellule végétale. Le procédé consiste à utiliser l'ADN exogène en combinaison avec une ou plusieurs protéines qui favorisent l'intégration de l'ADN exogène dans des cellules eucaryotes ciblées pour la transformation, les protéines provenant d'un gène chimère ou d'un ARN traduisible, pouvant s'exprimer dans la cellule eucaryote. Le procédé est applicable, en particulier, à des cellules végétales, et il permet une intégration stable d'un fragment d'ADN exogène délimité par des bordures d'ADN-T sous une forme intacte, en utilisant des protéines favorisant l'intégration provenant d'Agrobacterium. Des cultures, des tissus et des organismes entiers de nature transgénique, en particulier des plantes transgéniques, peuvent s'obtenir à partir de cellules transformées par le procédé de l'invention.

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