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Last Updated: April 26, 2024

Claims for Patent: 5,804,694


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Summary for Patent: 5,804,694
Title: Commercial production of .beta.-glucuronidase in plants
Abstract:A method for commercial production of GUS entails heterologous expression of the protein in plants, in native conformation, at an expression level such that avidin represents at least 0.1% of total extracted protein. A genetic map of the integration locus allows for the identification of the GUS-expressing plant. Genetic loci on a plant chromosome are revealed that support high levels of GUS expression and that can be used as a site of integration for high level expression of other genes of interest.
Inventor(s): Bruce; Wesley B. (Urbandale, IA), Hood; Elizabeth (Clive, IA), Peterson; David J. (Ames, IA), Register, III; James C. (Ames, IA), Witcher; Derrick (Urbandale, IA), Howard; John A. (West Des Moines, IA)
Assignee: ProdiGene, Inc. (College Station, TX)
Application Number:08/554,169
Patent Claims:1. A method of producing B-glucuronidase (GUS) in commercial quantities comprising the steps of (i) providing biomass from a plurality of plants, wherein at least some plants of said plurality express a DNA molecule comprised of (A) a heterologous nucleotide sequence coding for GUS and (B) a promoter operably linked to said nucleotide sequence to effect expression of GUS; and (ii) extracting GUS from said biomass wherein at least 0.1% of total extracted protein is GUS.

2. A method according to claim 1, wherein said biomass is comprised of seeds.

3. A method according to claim 1, wherein about 0.5% of total extracted protein in said biomass is GUS.

4. A method according to claim 3, wherein said biomass is comprised of seed.

5. A transgenic plant produced by a process comprising the steps of:

(a) providing cell or regenerable plant tissue that contains (i) a plant-compatible promoter, (ii) a signal sequence and (iii) a heterologous, GUS-encoding DNA sequence, wherein elements (i)-(iii) are operably linked,

(b) propagating a plurality of transgenic plants from said cell or said plant tissue, and then

(c) identifying from among said plurality a transgenic plant that expresses GUS such that at least 0.1% of total extracted protein is GUS.

6. A transgenic plant according to claim 5, wherein said heterologous nucleotide sequence encoding GUS incorporates plant-preferred codons.

7. A transgenic plant according to claim 5, wherein said DNA molecule further comprises an intron sequence or a peptide export signal sequence which modifies expression of said heterologous nucleotide sequence.

8. A transgenic plant according to claim 7, wherein said DNA molecule comprises said intron sequence.

9. A transgenic plant according to claim 7, wherein said DNA molecule comprises said peptide export signal sequence.

10. A transgenic plant according to claim 5, wherein said plant is a corn plant.

11. A transgenic plant according to claim 5, wherein said plant is strain 106155 and seeds of said strain are deposited under ATCC accession no. 97330.

12. A transgenic plant according to claim 5, wherein said plant is strain 106202 and seeds of said strain are deposited under ATCC accession no. 97329.

13. A transgenic plant according to claim 5, wherein about 0.5% of said total extracted protein is GUS.

14. A transgenic plant according to claim 5, wherein said promoter is the ubiquitin promoter.

15. A transgenic plant according to claim 5, wherein said promoter is a tissue-preferred promoter.

16. A transgenic plant according to claim 8, wherein said intron sequence is a plant ubiquitin intron sequence.

17. A transgenic plant according to claim 16, wherein said DNA molecule further comprises part of ubiquitin gene first exon.

18. A transgenic plant according to claim 9, wherein said peptide export signal sequence is a barley alpha amylase peptide export signal sequence.

19. Seed that is the product of a plant according to claim 5.

20. Seed that is the product of a plant according to claim 13.

Details for Patent 5,804,694

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 06/04/1986 ⤷  Try a Trial 2039-02-26
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2039-02-26
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2039-02-26
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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