Method for obtaining transgenic plants expressing a protein...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/82 (2006.01) A01H 5/00 (2006.01) C12N 9/02 (2006.01) C12N 9/16 (2006.01) C12N 9/18 (2006.01)

Patent

CA 2346153

The subject of the present invention is a method for obtaining transgenic plants expressing a protein with H2O2 producing activity, which comprises the following steps of: (a) transforming plant cells with Agrobacterium rhizogenes containing a vector carrying a gene encoding a protein producing H2O2 in a context which allows its expression in the plant; (b) selecting the transformants which contain and express this gene, by a peroxidase-based colorimetric test; (c) regenerating the plants from the roots selected and monitoring the expression of the plantlets obtained by a peroxidase-based colorimetric test; (d) sorting according to phenotype and optionally carrying out the molecular analysis of the progeny of the transgenic plants, allowing the selection or the confirmation of transgenic plants obtained containing only the transgene and not the T-DNA specific to A. rhizogenes.

La présente invention a pour objet un procédé pour l'obtention de plantes transgéniques exprimant une protéine à activité productrice de H¿2?O¿2?, qui comprend les étapes suivantes de: (a) transformation de cellules végétales par Agrobacterium rhizogenes contenant un vecteur porteur d'un gène codant pour une protéine productrice de H¿2?O¿2? dans un contexte qui permet son expression dans le végétal; (b) sélection des transformants qui contiennent et expriment ce gène par un test colorimétrique à la peroxydase; (c) régénération des plantes à partir des racines sélectionnées et contrôle de l'expression des plantules obtenues par un test colorimétrique à la peroxydase; (d) tri phénotypique et éventuellement analyse moléculaire de la descendance des plantes transgéniques, permettant la sélection ou la confirmation des plantes transgéniques obtenues contenant seulement le transgène et non l'ADN-T propre à A. rhizogenes. Application: obtention de plantes transgéniques.

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