Method for promoting homologous recombination
AssigneeKao Corporation
InventorsMayumi Wada, Shintaro Ryo
Life of this patent
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Abstract
A method of producing a transformant, which contains using an alga belonging to the genus Nannochloropsis as a host, wherein function of the following protein (A) or (B) of the alga is suppressed, inhibited or deleted: (A) a protein consisting of the amino acid sequence set forth in SEQ ID NO: 50; and (B) a DNA binding protein consisting of an amino acid sequence having 70% or more identity with the amino acid sequence of the protein (A).
Claims15
A method of producing a transformant, the method comprising:
- transforming a Nannochloropsis host with a polynucleotide and inserting the polynucleotide at a target site in the host's genomic DNA by homologous recombination;
- wherein, in the host:
- a gene encoding the following protein (A) or protein (B) has been deleted or inactivated prior to the transforming, thereby
- (i) reducing the host's frequency of non-homologous recombination, and
- (ii) increasing the host's frequency of homologous recombination;
- wherein, both (i) and (ii) are as compared to that of a Nannochloropsis alga that is the same as the host except that the gene encoding protein (A) or protein (B) is not deleted or inactivated:
- wherein protein (A) is:
- (A) a protein consisting of the amino acid sequence set forth in SEQ ID NO:
- 50;
- and
- wherein protein (B) is:
- (B) a DNA binding protein consisting of an amino acid sequence having 90% or more identity with the amino acid sequence of protein (A),
- and wherein protein (B) recognizes DNA ends generated by a DNA double strand break, and binds to the DNA ends recognized to recruit a DNA-dependent kinase to the DNA ends.
A method of producing a host, the method comprising deleting or inactivating a gene that encodes the following protein (A) or protein (B) in an alga belonging to the genus Nannochloropsis, thereby producing a host in which the host's frequency of non-homologous recombination is reduced and the host's frequency of homologous recombination is increased, both as compared to that of a Nannochloropsis alga that is the same as the host except that the gene encoding protein (A) or encoding protein (B) is not deleted or inactivated:
- wherein protein (A) is:
- (A) a protein consisting of the amino acid sequence set forth in SEQ ID NO:
- 50;
- and
- wherein protein (B) is:
- (B) a DNA binding protein consisting of an amino acid sequence having 90% or more identity with the amino acid sequence of protein (A),
- and wherein protein (B) recognizes DNA ends generated by a DNA double strand break, and binds to the DNA ends recognized to recruit a DNA-dependent kinase to the DNA ends.
An alga belonging to the genus Nannochloropsis, in which
- (a) a gene encoding the following protein (A) or protein (B) is deleted or inactivated;
- and
- (b) the alga's frequency of non-homologous recombination is reduced and the alga's frequency of homologous recombination is increased, both as compared to that of a Nannochloropsis alga that is the same except that the gene encoding protein (A) or the gene encoding protein (B) is not deleted or inactivated,
- wherein protein (A) is:
- (A) a protein consisting of the amino acid sequence set forth in SEQ ID NO:
- 50;
- and
- wherein protein (B) is:
- (B) a DNA binding protein consisting of an amino acid sequence having 90% or more identity with the amino acid sequence of protein (A), and wherein protein (B) is a protein that recognizes DNA ends generated by a DNA double strand break, and binds to the DNA ends recognized to recruit a DNA-dependent kinase to the DNA ends.
Dependencies are inferred from the claim text.