Methods and materials for making and using transgenic dicamba-degrading organisms

• US 8,629,323 B2
• Filed: 01/12/2012
• Issued: 01/14/2014
• Est. Priority Date: 04/04/1997
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Nebraska
• Original Assignee: Board of Regents of the University of Nebraska
• Inventors: Weeks, Donald P., Wang, Xiao-Zhuo, Herman, Patricia L.
• Primary Examiner(s): RAGHU, GANAPATHIRAM
• Application Number:
  US13/348,900
• Publication Number:
  US 20120151630A1
• Time in Patent Office: 733 Days
• Field of Search: 800/278, 800/288, 800/300, 435/189, 435/69.1, 435/320.1, 435/410, 435/413, 435/418, 435/419, 536/23.2, 536/23.7, 530/350
• US Class Current: 800/278
• CPC Class Codes: C07K 14/79 Transferrins, e.g. lactoferrins, ovotransferrins
• CPC Class Codes: C12N 15/8209 Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers
• CPC Class Codes: C12N 15/8259 Phytoremediation
• CPC Class Codes: C12N 15/8274 for herbicide resistance
• CPC Class Codes: C12N 9/00 Enzymes; Proenzymes; Compositions thereof preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113; Processes for preparing, activating, inhibiting, separating or purifying enzymes preparation of malt C12C1/00
• CPC Class Codes: C12N 9/0004 Oxidoreductases (1.)
• CPC Class Codes: C12N 9/0036 acting on NADH or NADPH (1.6)
• CPC Class Codes: C12N 9/0069 acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
• CPC Class Codes: C12N 9/1007 Methyltransferases (general) (2.1.1.)

Abstract

The invention provides isolated and at least partially-purified dicamba-degrading enzymes, isolated DNA molecules coding for dicamba-degrading enzymes, DNA constructs coding for dicamba-degrading enzymes, transgenic host cells comprising DNA coding for dicamba-degrading enzymes, and transgenic plants and plant parts comprising one or more cells comprising DNA coding for dicamba-degrading enzymes. Expression of the dicamba-degrading enzymes results in the production of dicamba-degrading organisms, including dicamba-tolerant plants. Finally, the invention provides a method of selecting transformed plants and plant cells based on dicamba tolerance and a method of selecting or screening transformed host cells, intact organisms and parts of organisms based on the fluorescence of 3,6-dichlorosalicylic acid produced as a result of dicamba degradation.

Claims

1. A method for producing a dicamba-degrading oxygenase that comprises the amino acid sequence of SEQ ID NO:

3, or that comprises an amino acid sequence that differs from SEQ ID NO;

3 by one substitution, an amino acid sequence that differs from SEQ ID NO;

3 by one addition, or an amino acid sequence that differs from SEQ ID NO;

3 by one substitution and by one addition, said method comprising culturing a host cell transformed with a replication or expression vector comprising DNA sequence encoding a dicamba-degrading oxygenase that comprises the amino acid sequence of SEQ ID NO;

3, or that comprises an amino acid sequence that differs from SEQ ID NO;

3 by one substitution, an amino acid sequence that differs from SEQ ID NO;

3 by one addition, or an amino acid sequence that differs from SEQ ID NO;

3 by one substitution and by one addition, under conditions effective to express said polypeptide, and recovering the polypeptide so expressed.

2. A seed containing a recombinant dicamba-degrading oxygenase comprising the amino acid sequence of SEQ ID NO:

3.

6. A seed of claim 2 wherein said oxygenase is produced by expression of DNA comprising SEQ ID NO:

2.

3. A seed containing a recombinant dicamba-degrading oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one substitution.

7. A seed of claim 3 wherein said oxygenase is produced by expression of a DNA encoding an oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one substitution.

4. A seed containing a recombinant dicamba-degrading oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one addition.

8. A seed of claim 4 wherein said oxygenase is produced by expression of a DNA encoding an oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one addition.

5. A seed containing a recombinant dicamba-degrading oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one substitution and by one addition.

9. A seed of claim 5 wherein said oxygenase is produced by expression of a DNA encoding an oxygenase comprising an amino acid sequence that differs from SEQ ID NO:

3 by one substitution and by one addition.