Alteration of amino acid compositions in seeds

• US 5,850,016 A
• Filed: 03/20/1996
• Issued: 12/15/1998
• Est. Priority Date: 03/20/1996
• Status: Expired due to Term

• Current Assignee: Pioneer Hi Bred International Inc (Corteva, Inc.)
• Original Assignee: Pioneer Hi Bred International Inc (Corteva, Inc.)
• Inventors: Hu, David, Coughlan, Sean, Jung, Rudolf, Hastings, Craig
• Primary Examiner(s): LeGuyader, John
• Assistant Examiner(s): McGarry, Sean
• Application Number:
  US08/618,911
• Time in Patent Office: 1,000 Days
• Field of Search: 435/6, 435/69.1, 435/172.1, 435/320.1, 435/410, 435/415, 536/23.1, 536/23.4, 536/23.6, 800/205, 800/250
• US Class Current: 800/287
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8253 Methionine or cysteine
• CPC Class Codes: C12N 15/8254 Tryptophan or lysine

Abstract

The present invention provides methods for increasing the level of preselected amino acids in seeds of plants, thereby enhancing the nutritional value of the seeds, by genetic modification. The present invention is particularly useful in increasing the methionine, lysine, and/or cysteine content in seeds of plants. Also provided, are isolated endogenous DNA molecules which encode soybean albumins. The present invention also provides an antibody which is capable of specifically binding to soybean albumins. The present invention further provides methods for isolating and purifying 2S albumins.

Claims

1. An isolated DNA molecule comprising a preselected DNA segment which encodes a protein having SEQ ID NOS:

2, 4 or 6.

2. An expression cassette comprising the DNA molecule of claim 1 operably linked to a promoter functional in a host plant cell.

3. The expression cassette of claim 2 wherein the promoter is a seed-specific promoter.

4. A fertile transformed plant containing the isolated preselected DNA segment of claim 2.

5. The plant of claim 4 which is a soybean plant.

6. A method of increasing the nutritional value of a seed comprising:

a) introducing into the cells of a plant the expression cassette of claim 3 to produce transformed plant cells;

b) regenerating a transformed plant from the transformed cells; and

c) isolating seeds from the regenerated transformed plant.

7. The method of claim 6 wherein the plant is a soybean plant.

8. The method of claim 6 wherein lysine, methionine, cysteine or combinations thereof are present in the seed in an amount greater than the amount in a seed of a corresponding untransformed plant.

9. The method of claim 8 wherein lysine is present in the seed in an amount at least about 5% greater than the amount in a seed of a corresponding untransformed plant.

10. The method of claim 8 wherein methionine and cysteine are present in the seed in an amount at least about 15% greater than the amount in a seed of a corresponding untransformed plant.

11. A seed produced by the method of claim 6.

12. A plant produced from the seed of claim 11.

13. An isolated DNA molecule comprising a preselected DNA segment comprising the sequence of SEQ ID NOS:

1, 3 or 5.

16. An expression cassette comprising the DNA molecule of claim 13 operably linked to a promoter functional in a host plant cell.

17. The expression cassette of claim 16 wherein the promoter is a seed-specific promoter.

18. A fertile transformed plant containing the preselected DNA segment of claim 13.

19. The plant of claim 18 wherein the preselected DNA segment encodes an endogenous polypeptide sequence.

20. The plant of claim 18 which is a soybean plant.

21. The plant of claim 20 wherein the preselected DNA segment encodes a soybean albumin.

22. A method of increasing the nutritional value of a seed comprising:

a) introducing into the cells of a plant the expression cassette of claim 16 to produce transformed plant cells;

b) regenerating a transformed plant from the transformed cells; and

c) isolating seeds from the regenerated transformed plant.

23. The method of claim 22 wherein the plant is a soybean plant.

24. The method of claim 23 wherein lysine, methionine, cysteine or combinations thereof are present in the seed in an amount greater than the amount in a seed of a corresponding untransformed plant.

25. The method of claim 24 wherein lysine is present in the seed in an amount at least about 5% greater than the amount in the seed of a corresponding untransformed plant.

26. The method of claim 24 wherein methionine and cysteine are present in the seed in an amount at least about 15% greater than the amount in the seed of a corresponding untransformed plant.

27. A seed produced by the method of claim 22.

28. A plant produced from the seed of claim 27.

14. A DNA molecule wherein SEQ ID NOS:

1 or 3 are modified by site-directed mutagenesis to encode for a protein with increased levels of lysine, methionine, cysteine or combinations thereof.

15. A DNA molecule wherein SEQ ID NOS:

1 or 3 are modified by site-directed mutagenesis to encode for a protein with decreased levels of asparagine and/or glutamine.