IP Library › Granted Patent US 10,745,711
Granted Patent B2
US 10,745,711 · App. 15/786,627 · Granted Aug 18, 2020

Soybean event DP-305423-1 and compositions and methods for the identification and/or detection thereof

Inventors: Anthony J Kinney (Wilmington, DE); Kent Brink (Newark, DE); Robert F. Cressman, Jr. (Wilmington, DE); Knut Meyer (Des Moines, IA); Kevin L. Stecca (Middletown, DE); Natalie N. Weber (Hockessin, DE); Cathy Xiaoyan Zhong (Wilmington, DE)
Assignee: E. I. DU PONT DE NEMOURS AND COMPANY
C12N15/8247A01H5/10C12N15/8274C12Q1/6895
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 10,745,711
App. No.
15/786,627
Granted
Aug 18, 2020
Kind
B2
Abstract

Compositions and methods related to transgenic high oleic acid/ALS inhibitor-tolerant soybean plants are provided. Specifically, the present invention provides soybean plants having a DP-305423-1 event which imparts a high oleic acid phenotype and tolerance to at least one ALS-inhibiting herbicide. The soybean plant harboring the DP-305423-1 event comprises genomic/transgene junctions having at least the polynucleotide sequence of SEQ ID NO:8, 9, 14, 15, 20, 21, 83 or 84. The characterization of the genomic insertion site of the DP-305423-1 event provides for an enhanced breeding efficiency and enables the use of molecular markers to track the transgene insert in the breeding populations and progeny thereof. Various methods and compositions for the identification, detection, and use of the soybean DP-305423-1 events are provided.

Claims (15)

1. A method of detecting the presence of DNA corresponding to the soybean event DP-305423-1 in a sample, the method comprising: (a) contacting a sample comprising soybean DNA with a primer set, wherein (I) a first DNA molecule of the primer set comprises at least 11 contiguous nucleotides or a full complement thereof of (i) nucleotides 18,652-31,579 of SEQ ID NO:5, (ii) nucleotides 12,164-14,494 of SEQ ID NO:6, (iii) nucleotides 5,751-7,813 of SEQ ID NO:7, or (iv) nucleotides 2899-7909 of SEQ ID NO:82 and (II) a second DNA molecule of the primer set binds to the sample comprising soybean DNA to function to produce a DNA amplicon molecule for soybean event DP-305423-1 when used in a nucleic acid amplification reaction comprising the first DNA molecule and DNA from soybean event DP-305423-1; (b) performing a nucleic acid amplification reaction, thereby producing the DNA amplicon molecule; and (c) detecting the DNA amplicon molecule.

2. The method of claim 1 , wherein the DNA amplicon molecule comprises SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof.

3. The method of claim 1 , wherein the DNA amplicon molecule is detected in step (c) by hybridizing the DNA amplicon molecule to a probe which binds to SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof, under stringent hybridization conditions.

4. A method of detecting the presence of a nucleic acid corresponding to event DP-305423-1 in a sample, the method comprising: (a) contacting a sample comprising soybean DNA comprising event DP-305423-1 with a first primer and a second primer to function when used in a nucleic acid amplification reaction to amplify a nucleotide sequence in the soybean DNA, the nucleotide sequence comprising at least one of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof; (b) amplifying the nucleotide sequence to produce a polynucleotide comprising at least one of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof; and (c) detecting the polynucleotide amplified in step (b), thereby detecting the presence of a nucleic acid corresponding to event DP-305423-1 in the sample.

5. The method of claim 4 , wherein the polynucleotide is detected in step (c) by hybridizing the polynucleotide to a probe which binds to SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof, under stringent hybridization conditions.

6. A method of detecting the presence of a DNA molecule from soybean event DP-305423-1 comprising: (a) contacting a DNA sample from a soybean plant comprising soybean event DP-305423-1 with a pair of DNA molecules consisting of a first DNA molecule and a second DNA molecule different from the first DNA molecule, wherein the DNA molecules have a nucleotide sequence of sufficient length of contiguous nucleotides of SEQ ID NO: 5, 6, 7, or 82, or a complete complement thereof, and wherein the first DNA molecule resides in a transgene insert DNA sequence of SEQ ID NO: 5, 6, 7, or 82, and the second DNA molecule resides in the corresponding soy genomic DNA sequence of SEQ ID NO: 5, 6, 7, or 82; (b) performing an amplification reaction sufficient to produce a DNA amplicon; and (c) detecting the DNA amplicon, wherein detection of the DNA amplicon is diagnostic for the presence of the DNA molecule from soybean event DP-305423-1 in the DNA sample.

7. The method of claim 6 , wherein the DNA amplicon molecule is detected in step (c) by hybridizing the DNA amplicon molecule to a probe which binds to SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof, under stringent hybridization conditions.

8. The method of claim 6 wherein the amplicon comprises at least 15 contiguous nucleotides of SEQ ID NO: 8, 9, 14, 15, 20, 21, 83 or 84, or a complete complement thereof.

9. A method of detecting the presence of DNA corresponding to the soybean DP-305423-1 event in a sample, the method comprising: (a) contacting the sample comprising soybean DNA with a polynucleotide probe that hybridizes under stringent hybridization conditions with DNA from soybean DP-305423-1 event and does not hybridize under said stringent hybridization conditions with a non-DP-305423-1 soybean plant DNA; (b) subjecting the sample and probe to stringent hybridization conditions; and (c) detecting hybridization of the probe to the DNA; wherein detection of hybridization indicates the presence of the DP-305423-1 event.

10. The method of claim 9 , wherein the probe comprises the 5′ junction sequence of event DP-305423-1.

11. The method of claim 9 , wherein the probe comprises the 3′ junction sequence of event DP-305423-1.

12. The method of claim 9 , wherein the probe comprises SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof.

13. The method of claim 9 , wherein the hybridization is detected in step (c) by hybridizing the DNA amplicon molecule to a probe which binds to SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, or SEQ ID NO:88, or a full complement thereof.

14. The method of claim 9 , wherein the polynucleotide probe binds to a sequence within nucleotides 18,652-31,579 of SEQ ID NO:5, nucleotides 12,164-14,494 of SEQ ID NO:6, nucleotides 5,751-7,813 of SEQ ID NO:7 or nucleotides 2,899-7,909 of SEQ ID NO:82.

15. The method of claim 9 , wherein the polynucleotide probe binds to a sequence within nucleotides 18,652-31,579 of SEQ ID NO:5.

Assignments (1)
NUNC PRO TUNC ASSIGNMENT Recorded Nov 29, 2022
From: E.I. DU PONT DE NEMOURS AND COMPANY
To: CORTEVA AGRISCIENCE LLC
Reel/Frame 063141/0155 →
Continuity (5)
Division 14053059 · Oct 14, 2013
Continuation 11927884 · Oct 30, 2007
Provisional Application 60942676 · Jun 8, 2007
Provisional Application 60863721 · Oct 31, 2006
Related Publication 20180030464A1 · Feb 1, 2018