Patent filed by Japan Tobacco – Actual filed independent claims

Patent filed by Japan Tobacco, Actual filed independent claims

EP 927765 A1

Claim 1A method for selecting transformed cells which comprises: A) culturing cells originating from rice tissue in a selective medium containing paromomycin, after transferring at least a desired structural gene and a paromomycin resistance gene into the cells, and
B) selecting transformed cells.
Claim 7A transformed rice cell which is selected from cells originating from rice tissue by culturing the cells in a selective medium containing paromomycin after at least a desired structural gene and a paromomycin resistance gene were transferred into the cells.
Claim 8A method for producing a rice transformant transformed by a desired gene comprising:
A) providing a strain belonging to the genus Agrobacterium which has a plasmid containing a paromomycin resistance gene and a desired gene, in the T-DNA region in said plasmid, in such a way as to allow expression of each of said genes;
B) providing cells originating from rice tissue;
C) inoculating the above cells originating from rice with the above strain belonging to the genus Agrobacterium;
D) culturing the inoculated cells in a culture medium for plant cells containing paromomycin at such a concentration that will not allow cells other than those having been transformed by the paromomycin resistance gene to survive in said medium (hereinafter referred to as a selective medium), and selecting paromomycin-resistant calluses;
E) if necessary, repeating the selection step d) once or more by using tissue cells obtained from the selected calluses; and
F) culturing the selected calluses in a medium appropriate for plant regeneration to give a completely regenerated plant.