Enhanced zein reduction in transgenic corn seed

A method for generating corn seeds having at least one of enhanced lysine, enhanced oil and enhanced tryptophan comprising the steps of:

• a) transforming a plant cell with the recombinant DNA construct for suppression of a plurality of maize zein target genes, comprising in 5′ to 3′ order a promoter element operably linked to a plurality of anti-sense-oriented DNA elements from a plurality of maize zein target genes, the anti-sense-oriented DNA elements designated in 5′ to 3′ order element 1, element 2, and a plurality of sense-oriented DNA elements from a plurality of maize zein target genes, designed in 5′ to 3′ order element 3, element 4, wherein the anti-sense-oriented DNA element 2 has at least a portion of its sequence at its 3′ end that is not complementary either to itself or to the sense-oriented DNA element 3, and wherein sense-oriented RNA transcribed by the sense-oriented DNA elements is complementary to the 5′-most end of anti-sense-oriented RNA transcribed by the anti-sense-oriented DNA elements;
• and b) regenerating the plant cell into a fertile transgenic plant, wherein the plant contains the construct in its genome;
• and c) harvesting seed from the plant, wherein the seed has at least one of enhanced lysine, enhanced oil and enhanced tryptophan relative to seed of the same variety not transformed with the construct.

A method for generating corn seeds having at least one of enhanced lysine, enhanced oil and enhanced tryptophan comprising the steps of:

• transforming a plant cell with the recombinant DNA construct for suppression of more than one maize zein target gene that comprises in 5′ to 3′ order a promoter element operably linked to more than one anti-sense-oriented DNA element from more than one maize zein target gene, the anti-sense-oriented DNA elements designed in 5′ to 3′ order element 1, element 2, and at least one sense-oriented DNA element from at least one maize zein target gene, designated element 3, wherein the anti-sense-oriented DNA element 2 has at least a portion of its sequence at its 3′ end that is not complementary either to itself or to the sense-oriented DNA element 3, and sense-oriented RNA transcribed by the sense-oriented DNA element is complementary to the 5′-most end of anti-sense-oriented RNA transcribed by the anti-sense-oriented DNA element(s);
• and regenerating the plant cell into a fertile transgenic plant, wherein the plant contains the construct in its genome;
• and harvesting seed from the plant, wherein the seed has at least one of enhanced lysine, enhanced oil and enhanced tryptophan relative to seed of the same variety not transformed with the construct.

A transgenic plant comprising within its genome a DNA construct of for suppression of a plurality of maize zein target genes, comprising in 5′ to 3′ order a promoter element operably linked to a plurality of anti-sense-oriented DNA elements from a plurality of maize zein target genes, the anti-sense-oriented DNA elements designated in 5′ to 3′ order element 1, element 2, and a plurality of sense-oriented DNA elements from a plurality of maize zein target genes, designed in 5′ to 3′ order element 3, element 4, wherein the anti-sense-oriented DNA element 2 has at least a portion of its sequence at its 3′ end that is not complementary either to itself or to the sense-oriented DNA element 3, and wherein sense-oriented RNA transcribed by the sense-oriented DNA elements is complementary to the 5′-most end of anti-sense-oriented RNA transcribed by the anti-sense-oriented DNA elements.

A transgenic plant comprising within its genome a DNA construct for suppression of more than one maize zein target gene that comprises in 5′ to 3′ order a promoter element operably linked to more than one anti-sense-oriented DNA element from more than one maize zein target gene, the anti-sense-oriented DNA elements designed in 5′ to 3′ order element 1, element 2, and at least one sense-oriented DNA element from at least one maize zein target gene, designated element 3, wherein the anti-sense-oriented DNA element 2 has at least a portion of its sequence at its 3′ end that is not complementary either to itself or to the sense-oriented DNA element 3, and sense-oriented RNA transcribed by the sense oriented DNA element is complementary to the 5′-most end of anti-sense-oriented RNA transcribed by the anti-sense-oriented DNA element(s).