Modified vird2 protein and its use in improved gene transfer

Details Modified vird2 protein and its use in improved gene transfer Modified vird2 protein and its use in improved gene transfer

C - Chemistry, Metallurgy

12

N

C12N 15/82 (2006.01) A01H 5/00 (2006.01) C07K 14/195 (2006.01)

Patent

CA 2652266

There is provided a method for Agrobacterium-mediated gene transformation of a host cell. The method uses an Agrobacterium which expresses a VirD2 protein resistant to cleavage by a caspase. The VirD2 protein is modified by replacement of an aspartic acid residue with an alternative amino acid at one or more of the caspase cleavage sites within the protein amino acid sequence. This method improves the low efficiency of Agrobacterium-mediated gene transfer in certain plant cells and in animal cells caused by caspase cleavage of the VirD2 protein. The modified VirD2 protein, a gene for its expression and a method of producing a protein of interest by Agrobacterium-mediated transformation of a host cell are also described.

L'invention concerne un procédé de transformation génique d'une cellule hôte au moyen d'Agrobacterium. Le procédé utilise une bactérie Agrobacterium exprimant une protéine VirD2 résistante au clivage par une caspase. La protéine VirD2 est modifiée par remplacement d'un résidu acide aspartique par un autre acide aminé au niveau d'un ou plusieurs des sites de clivage par caspase à l'intérieur de la séquence d'acides aminés de la protéine. Ce procédé améliore la faible efficacité du transfert de gène médié par Agrobacterium chez certaines cellules végétales et dans les cellules animales, ladite faible efficacité étant due au clivage de la protéine VirD2 par une caspase. L'invention concerne également la protéine VirD2 modifiée, un gène permettant d'exprimer ladite protéine et un procédé de production d'une protéine d'intérêt par transformation d'une cellule hôte au moyen d'une bactérie Agrobacterium.

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