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Last Updated: January 29, 2022

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Claims for Patent: 7,064,248

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Summary for Patent: 7,064,248
Title:Method of preparing fertile transgenic corn plants by microprojectile bombardment
Abstract: A method of preparing fertile transgenic Zea mays (corn) plants which stably express DNA encoding a Bacillus thuringiensis endotoxin, so as to impart insect resistance to the fertile transgenic plants is taught. The method comprises bombarding regenerable corn cells with DNA-coated microprojectiles, wherein the DNA is adjusted to be more efficiently expressed in corn than the native B. thuringiensis sequence, and regenerating a fertile transgenic plants from the transformed cells.
Inventor(s): Lundquist; Ronald C. (Minnetonka, MN), Walters; David A. (Bloomington, MN), Kirihara; Julie A. (Bloomington, MN)
Assignee: DeKalb Genetics Corp. (DeKalb, IL)
Application Number:09/818,921
Patent Claims:1. A process for producing a fertile transgenic Zea mays plant comprising the steps of (i) bombarding intact regenerable Zea mays cells with DNA-coated microprojectiles; wherein said DNA comprises a preselected DNA sequence encoding a Bacillus thuringiensis endotoxin, wherein the preselected DNA sequence is adjusted to be more efficiently expressed in Zea mays than the native B. thuringiensis DNA sequence encoding said endotoxin; (ii) identifying a population of transformed cells comprising said preselected DNA sequence; and (iii) regenerating a fertile transgenic plant therefrom, wherein said preselected DNA sequence is expressed so as to impart insect resistance to said transgenic plant and is heritable.

2. The process of claim 1, wherein the preselected DNA sequence encoding said endotoxin comprises an increased number of maize preferred codons compared to native Bacillus thuringiensis endotoxin.

3. The process of claim 1, wherein the preselected DNA sequence further comprises a protease inhibitor.

4. A population of plants obtained by breeding the transgenic plants of claim 1 wherein the preselected DNA sequence from said transgenic plant is transmitted by Mendelian inheritance through both male and female parent plants to the population of plants.

5. The process of claim 1, wherein the preselected DNA sequence further comprises a selectable marker gene or a reporter gene.

6. The process of claim 5, wherein the selectable marker gene confers resistance or tolerance to a compound selected from the group consisting of hygromycin, sethoxydim, haloxyfop, glyphosate, methotrexate, imidazoline, sufolnylurea, triazolopyrimidine, s-triazine, bromoxynil, phosphinotliricin, kanamycin, G418, 2,2-dichloropropionic acid and neomycin.

7. The process of claim 6, wherein the compound is phosphinothricin.

8. The process of claim 6, wherein the compound is kanamycin.

9. The process of claim 6, wherein the compound is hygromycin.

10. The process of claim 5, wherein the DNA sequence encoding the Bacillus thuringiensis endotoxin is fused in frame with said selectable marker or reporter gene.

11. The process of claim 5, wherein the preselected DNA sequence encoding said endotoxin comprises an increased number of maize preferred codons compared to native Bacillus thuringiensis endotoxin.

12. The process of claim 1 or 5, wherein the fertile transgenic Zea mays plant is generated from transformed embryogenic tissue.

13. The process of claim 12, wherein the cells are derived from immature embryos.

14. The process of claim 1 or 5, further comprising obtaining transgenic insect resistant progeny plants of subsequent generations from said fertile transgenic plant.

15. The process of claim 14, further comprising obtaining seed from one of said progeny plants.

16. The process of claim 1 or 5, wherein the preselected DNA sequence encodes the HD73 endotoxin of Bacillus thuringiensis.

17. The process of claim 16, wherein the preselected DNA sequence encoding said endotoxin comprises an increased number of maize preferred codons compared to native Bacillus thuringiensis endotoxin.

18. The process of claim 1 or 5, wherein the preselected DNA sequence encodes the HD1 endotoxin of Bacillus thuringiensis.

19. The process of claim 18, wherein the preselected DNA sequence encoding said endotoxin comprises an increased number of maize preferred codons compared to native Bacillus thuringiensis endotoxin.

20. The process of claim 1 or 5, wherein the preselected DNA sequence encodes a truncated endotoxin of Bacillus thuringiensis.

21. The process of claim 20, wherein the Bacillus thuringiensis endotoxin comprises about the N-terminal 50% of the endotoxin.

22. The process of claim 1 or 5, wherein the preselected DNA sequence comprises an operably linked promoter.

23. The process of claim 5, wherein the preselected DNA sequence further comprises a promoter operably linked to said DNA sequence encoding said endotoxin and a promoter operably linked to said selectable marker gene.

24. The process of claim 22, wherein the preselected DNA sequence further comprises the maize AdhIS first intron or the maize Shrunken-2 first intron positioned between the promoter and the preselected DNA sequence encoding said endotoxin.

25. The process of claim 22, wherein the preselected DNA sequence further comprises a manopine synthase promoter, a nopaline synthase promoter or an octopine synthase promoter operably linked to said preselected DNA sequence.

26. The process of claim 22, wherein the promoter is the CaMV 35S or 19S promoter.

Details for Patent 7,064,248

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 1986-06-04 ⤷  Try it Free 2010-01-22
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try it Free 2010-01-22
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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