DP-023211-2 Corn Detection via Flanking Sequence Primers
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Solution Overview
Problem
Current methods for detecting transgenic insect-resistant corn plants, such as those containing the DP-023211-2 event, face challenges in distinguishing between different events and progeny due to interchangeable coding regions and lack of specificity in DNA constructs, particularly when using PCR or DNA hybridization methods.
Innovation Solution
The development of a DNA construct and associated detection methods for the DP-023211-2 event in corn plants, utilizing specific primers and probes that target unique 5′ and 3′ flanking sequences, allowing for the identification of the event in progeny and biological samples through nucleic acid amplification and hybridization reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If PCR or DNA hybridization methods are used to detect transgenic corn plants, then detection capability is achieved, but specificity to distinguish between different events is lost due to interchangeable coding regions
Solution Approach 1:
The detection method is divided into multiple independent components: a first primer binding to the first flanking sequence, a second primer binding to the second flanking sequence, and optionally a probe binding to the coding region. This segmentation allows each component to contribute to overall specificity, enabling distinction between different transgenic events even when coding regions are interchangeable.
Solution Approach 2:
The invention utilizes unique local characteristics of the flanking sequences (which differ between events) rather than relying on the interchangeable coding regions. By designing primers that bind to these unique flanking sequences, the method achieves event-specific detection despite the similarity or interchangeability of the coding regions themselves.
2Ease of manufacture
If detection methods focus on frequently used genetic elements like promoters and terminators, then detection is simplified, but ability to discriminate between different events is reduced
Solution Approach 1:
Instead of targeting common genetic elements like promoters and terminators that are shared across different events, the invention targets the unique flanking sequences that are specific to each event. This approach maintains relative simplicity while achieving the ability to discriminate between different events, as the flanking sequences serve as unique identifiers for each transgenic event.
Solution Approach 2:
The method incorporates knowledge of the specific flanking sequences associated with each transgenic event into the primer design stage. This preliminary customization of primers to match event-specific flanking sequences enables both simplicity in execution and precision in discrimination, as the specificity is built into the detection method before implementation.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables precise identification of the DP-023211-2 event in corn plants and progeny, providing a reliable method for distinguishing it from non-transgenic material and other transgenic events, ensuring accurate segregation and quality assessment of transgenic plant material.
Implementation Method 1
detect the presence of a transgene by a nucleic acid detection method by, e.g., a polymerase chain reaction (PCR)
Implementation Method 2
detect the presence of a transgene by a nucleic acid detection method by, e.g., a polymerase chain reaction (PCR) or DNA hybridization using nucleic acid probes
Data Source
AI summary
Embodiments disclosed herein relate to the field of plant molecular biology, specifically to DNA constructs for conferring insect resistance to a plant. Embodiments disclosed herein relate to insect resistant corn plant containing event DP-023211-2, and to assays for detecting the presence of event DP-023211-2 in samples and compositions thereof.