Transformation of eukaryotic cells by mobilizable plasmids

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/82 (2006.01) A01H 5/00 (2006.01) C12N 15/74 (2006.01) C12N 15/80 (2006.01) C12N 15/81 (2006.01)

Patent

CA 2401909

The invention relates to the transfer of genetic material to eukaryotic cells by means of a process resembling conjugation, in particular by a system partially based on Agrobacterium tumefaciens-like transfer systems. In particular transfer of genetic material into plant cells is disclosed using mobilisable, but non-conjugative plasmids by means of an Agrobacterium virulence system. The invention provides a method for transferring genetic material which is not a typical T-DNA surrounded by Agrobacterium T-borders from an Agrobacterium virulence system to an eukaryotic host cell, comprising providing said genetic material on a mobilisable plasmid, capable of forming a relaxosome, bringing said mobilisable plasmid in an Agrobacterium having at least the activity of the transfer genes of Agrobacterium not present on said mobilisable plasmid, whereby the necessary gene products providing the same or similar activity as a functional VirB operon are also present and cocultivating said Agrobacterium with said eukaryotic host cell.

L'invention concerne le transfert de matériau génétique à des cellules eucaryotes par un processus qui ressemble à la conjugaison, notamment selon un système reposant en partie sur les systèmes du type Agrobacterium tumefaciens. L'invention concerne en particulier le transfert de matériau génétique dans des cellules de plantes, par le biais de plasmides mobilisables non conjugatifs, sur la base d'un système à virulence d'Agrobacterium. L'invention concerne par ailleurs un procédé de transfert de matériau génétique différent de l'ADN typique, cadré par les limites T d'Agrobacteirum, depuis un système à virulence d'Agrobacterium vers une cellule hôte eucaryote, qui comprend les étapes suivantes: fourniture du matériau génétique sur un plasmide mobilisable, capable de former un relaxosome, introduction du plasmide dans un Agrobacterium ayant au moins l'activité des gènes de transfert de l'Agrobacterium ne se trouvant pas sur le plasmide en question, moyennant quoi les produits géniques nécessaires assurant une activité équivalente ou similaire à celle d'un opéron VirB fonctionnel sont également présents, et coculture de l'Agrobacterium considéré avec la cellule eucaryote.

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