HOMOLOGICAL RECOMBINATION MEDIATED BY ZINC FINGER NUCLEASES

摘要

The invention relates to a method for introducing of exogenous sequence into plant cell genome, wherein the method comprises the following stages: a) contacting of cell with the vector of donor DNA, containing the first, the second, and the fifth DNA sequences, where the mentioned the fifth sequence contains exogenous sequence of vector DNA which is entitled to introduction, and located between the mentioned the first and the second sequences of donor DNA vector; where the first sequence has at least 90-95% homology with the third sequence in plant cell genome; the second sequence has at least 90-95% homology with the forth sequence in plant cell genome; where the third and the forth sequences in plant cell genome represent sequences of chromosomal DNA, and the neighbor ends of the mentioned the third and the forth sequences are separated with at least one nucleotide pair; and (b) expression of one or more nucleases in the cell, where each of the mentioned one or more nucleases represents the pair of fused proteins, where each protein represent fusion between restriction endonuclease Fok I cleavage domain and constructed binding domain of zinc finger type, and where the mentioned one of more nucleases cleave the chromosome DNA in one or more corresponding target sequences between the third and the forth sequences, where the mentioned one or more nucleases cleave the chromosome DNA in position at 0.4-3 thousands base pairs from the mentioned the third or the forth sequences in plant cell genome; in such a way that cleavage of chromosome DNA on the stage (b) stimulates integration of the fifth DNA sequence by homological recombination.