Polynucleotides for mutagenesis in fungus comprising a functional gene in magnaporthe and an impala transposon
AssigneeAventis CropScience GmbH
InventorsMarie-Claire Grosjean-Cournoyer, Francois Villalba, Marc-Henri Lebrun, Marie-Josee Daboussi
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Abstract
The invention concerns a new tool for efficient mutagenesis enabling the generation of a collection of mutants in fungi by random insertion of a characterized Fusarium oxysporum Impala transposon in the genome of said fungi. The invention also concerns the resulting mutants.
Claims18
An isolated polynucleotide, characterized in that it comprises a marker gene which is inactivated by the insertion of an Impala transposon, said marker gene comprising, in the direction of transcription, a promoter regulatory sequence which is functional in Magnaporthe grisea and which is functionally linked to the coding sequence of said marker gene.
The polynucleotide as claimed in 1, characterized in that the Impala transposon is integrated into the promoter regulatory sequence of the polynucleotide as claimed in the invention.
An insertion mutant of filamentous fungi chosen from the fungi of the Magnaporthe or Penicillium genera, into the genome of which is integrated the Impala transposon.
A method for preparing insertion mutants of fungi, characterized in that it comprises the following steps:
- (a) said fungus is transformed with a polynucleotide comprising a marker gene which has been inactivated by the insertion of a defective Impala transposon under conditions which allow the excision of the Impala transposon of said marker gene and its reinsertion into the genome of the fungus;
- (b) the insertion mutants expressing the marker gene are identified.
A method for preparing insertion mutants of fungi, characterized in that it comprises the following steps:
- (a) said fungus is transformed with a polynucleotide comprising a marker gene which has been inactivated by the insertion of a defective Impala transposon;
- (b) defective Impala transposon is mobilized using a transposase, the expression of which is controlled, under conditions which allow the excision of the defective Impala transposon, its reinsertion and its stabilization in the genome of the fungus;
- (c) the insertion mutants expressing the marker gene are identified.
Dependencies are inferred from the claim text.