Methods and constructs for producing transgenic plants and methods of recovering expressed proteins

• US 8,252,552 B2
• Filed: 11/09/2006
• Issued: 08/28/2012
• Est. Priority Date: 04/11/2000
• Status: Expired due to Fees

• Current Assignee: Texas A&M University System
• Original Assignee: Texas A&M University System
• Inventors: Mirkov, T. Erik, Monclin, Jean P., Barrilleaux, Adam, Irvine, James E., Moonan, Francis
• Primary Examiner(s): DESAI, ANAND U
• Application Number:
  US11/558,247
• Publication Number:
  US 20070130655A1
• Time in Patent Office: 2,119 Days
• Field of Search: None
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 15/8257 for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 15/8279 for biotic stress resistance, pathogen resistance, disease resistance

Abstract

This invention relates to a regulatory element useful for genetically engineering sugarcane or other monocots, to the transformation of the monocots with the regulatory element so that they produce a desired product, and to the regeneration of the monocots transformed with the regulatory element. In particular the present invention provides a nucleic acid encoding the promoter of a sugarcane proline rich protein as shown in Seq ID #3.

Claims

1. A method of refining sugarcane to extract a transgenic protein comprising the steps of:

a. producing said transgenic protein in a transformed sugarcane plant;

b. extracting juice containing said transgenic protein from stalks of said transgenic sugarcane plant;

c. increasing the pH of the juice up to no more than pH 7;

d. cleaning said juice to remove particulate matter; and

e. transmitting said juice through at least one membrane in order to produce two fractions, one of said fractions containing said transgenic protein of interest.

2. The method of claim 1, wherein said transgenic protein is selected from the group consisting of a lectin, an enzyme, a vaccine, a bacterial lytic peptide, a bacterial lytic protein, an antimicrobial peptide, an antimicrobial peptide protein, an antiviral peptide, an antiviral protein, an insecticidal peptide, an insecticidal protein, a therapeutic peptide, and a therapeutic protein.

3. The method of claim 1, wherein said cleaning involves passing said juice through at least one screen.

4. The method of claim 3, wherein said screen removes particulates larger than about 100 to 150 microns in size.

5. The method of claim 1, wherein said membrane removes particulates larger than about 0.05 to 0.2 microns.

6. The method of claim 1, wherein said membrane removes particulates larger than about 150 kDa molecular weight.

7. A method of producing a peptide or protein comprising the steps of:

a. producing a transgenic sugarcane plant transformed with a gene coding for said peptide or protein;

b. harvesting cane stalks from said transgenic sugarcane plant;

c. extracting sugarcane juice;

d. increasing the pH of the juice up to no more than pH 7;

e. cleaning said juice to remove particulate matter; and

f. transmitting said juice through at least one membrane in order to produce two fractions, one of said fractions containing said transgenic protein of interest.

8. The method of claim 7, wherein said transgenic peptide or protein is selected from the group consisting of a lectin, an enzyme, a vaccine, a bacterial lytic peptide, a bacterial lytic protein, an antimicrobial peptide, an antimicrobial protein, an antiviral peptide, an antiviral protein, an insecticidal peptide, an insecticidal protein, a therapeutic peptide, and a therapeutic protein.

9. The method of claim 8, wherein said sugarcane juice is passed through at least one screen.

10. The method of claim 9, wherein said screen removes particulates larger than about 150 microns in size.

11. The method of claim 9, wherein said membrane removes particulates larger than about 0.05 to 0.2 microns.

12. The method of claim 9, wherein said membrane removes particulates larger than about 150 kDa molecular weight.