Method for the mutagenesis of nucleotide sequences in plants, algae or fungi

Abstract

The invention relates to a method for the mutagenesis of eukaryotic nucleotide sequences, preferably from plants, algae and/or fungi and a method for the production of genetically modified eukaryotic cells. The subject of the invention is the genetically altered nucleotide sequences of the type disclosed, vectors containing such nucleotide sequences, as well as genetically altered eukaryotic cells, tissue and/or parts of plants, algae and/or fungi and/or regenerated whole plants and the use thereof.

1 .- 10 . (canceled)  
     
     
         11 . A genetically-modified eukaryotic nucleotide sequence generated by a method comprising: 
 transferring a genomic nucleotide sequence or a cDNA sequence from a eukarmotic organism to a microorganism,    genetically modifying the genomic nucleotide sequence or the cDNA sequence in the microorganism by sequence-independent insertion mutagenesis via cointegrate formation dung conjugation between two microorganisms; and    isolating the genetically modified, genomic nucleotide sequence or cDNA sequence from the microorganism.    
     
     
         12 .- 24 . (canceled)  
     
     
         25 . The sequence of  claim 11 , wherein the genomic nucleotide sequence or the cDNA sequence originates from plants, algae or fungi.  
     
     
         26 . The sequence of  claim 11 , wherein the mutagenesis is effected with high efficiency with a relative frequency post-selection in a range of from approximately 90 to 100%.  
     
     
         27 . The sequence of  claim 11 , wherein a vector comprising a nucleotide sequence and additional functional nucleotide sequences is transferred to said microorganism.  
     
     
         28 . The sequence of  claim 11 , wherein a population of different eukaryotic nucleotide sequences is generated by sequence-independent mutagenesis based on a selected region of a eukaryotic nucleotide sequence.  
     
     
         29 . The sequence of  claim 11 , wherein the mutagenesis is effected by insertion of a heterologous nucleotide sequence.  
     
     
         30 . The sequence of  claim 29 , wherein the heterologous nucleotide sequence is a transposon.  
     
     
         31 . The sequence of  claim 11 , wherein the microorganism comprises characteristics for transposition and conjugation.  
     
     
         32 . The sequence of  claim 11 , wherein the microorganism is a bacterium of the family Enterobacteriaceae or Bacillaceae.  
     
     
         33 . A vector comprising the genetically-modified eukaryotic nucleotide sequence of  claim 11 .  
     
     
         34 . The vector of  claim 33 , further comprising one or more of the characteristics shown in  FIG. 1 ,  FIG. 2 ,  FIG. 5  or  FIG. 6 .  
     
     
         35 . The vector of  claim 33 , further comprising one or more additional functional nucleotide sequences.  
     
     
         36 . The vector of  claim 33 , further comprising one or more of the characteristics shown in  FIG. 3 .  
     
     
         37 . Use of the vector of  claim 33  in an organism.