Methods for altering the content of amino acids in Plants by mutations of displacement of Frame

摘要

Claim 1: a method for altering the content of amino acids of a polypeptide.Evaluate, characterized in that it comprises two or more Reading Frames in a Nucleic Acid coding for the polypeptide.Identify a reading frame encoding an Amino Acid Sequence that has a desired content of amino acidsAnd introduce a mutation of displacement of frame in the Nucleic Acid such that when the Nucleic Acid Sequence is expressed in a cell,It expresses the polypeptide which has the desired content of amino acids.Claim: a method to generate a part of Plant, Plant, or Plant Cell with altered levels of amino acidsWherein the method comprises: (a) contacting a Plant, PlantOr Plant Cell with a First Cutting endonuclease unconventional for a sequence in an endogenous Gene in the Plant, part of the plant or Plant Cell.Such that the first unconventional Cutting endonuclease generates a double Strand cut in or near the sequence to which it is addressed; (b) select a Plant, PlantOr Plant cell containing a mutation in the Gene displacement framework; (c) contacting a plantPart of the plant or Plant Cell with a Second conventional Cutting endonuclease not addressed to a sequence in the same Gene, which is directed to the first Cutting endonuclease not convincedNal, such that the second endonuclease generates an unconventional Cut cut in or near the double Strand sequence to which it is addressed; and (d) select a Plant, PlantOr Plant Cell that contains a Second mutation in the endogenous Gene.Claim 62: a Plant, part of the plant or Plant Cell with altered levels of amino acidsCharacterized in that the plant contains a mutation of displacement of frame in an exon of a Gene selected.Claim 67: The Plant, part of Plant, Plant Cell or the claim 62, wherein part of the Plant, Plant Cell, plant is a plant or Plant part.Plant Cell or cassava, Wheat, alfalfa, Oats, maize, Rice, Sorghum, potato, tomato, soybean or canola.