Maize Event DP-915635-4 Detection via Flanking DNA
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Solution Overview
Problem
Existing methods for detecting transgenic insect-resistant corn events, such as DP-915635-4, are inadequate for distinguishing between different events due to interchangeable coding regions and lack of specificity, especially when using DNA constructs with similar flanking DNA sequences.
Innovation Solution
Development of DNA constructs and detection methods utilizing specific primers and probes that recognize unique 5′ and/or 3′ flanking sequences of DP-915635-4, enabling accurate identification through PCR and hybridization techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional transgene detection methods (PCR or DNA hybridization) are used, then detection of transgene presence is achieved, but discrimination between different transgenic events is not possible due to interchangeable coding regions and similar flanking DNA sequences
Solution Approach 1:
The invention extracts and targets only the unique flanking DNA sequences adjacent to the inserted transgene, rather than detecting the interchangeable coding regions. By designing detection primers and probes that specifically bind to these unique flanking sequences, the method achieves event-specific discrimination while ignoring the variable coding portions of different transgenes.
Solution Approach 2:
The detection method focuses on the local unique characteristics of flanking DNA sequences rather than the overall transgene structure. Each transgenic event has distinct flanking sequences at its insertion site, and the invention exploits this local uniqueness to differentiate between events, even when their coding regions are identical or highly similar.
2Measurement precision
If detection methods target frequently used genetic elements (promoters, terminators, marker genes), then detection sensitivity is improved, but ability to discriminate between different events is lost
Solution Approach 1:
The invention extracts and targets only the unique flanking DNA sequences adjacent to the inserted transgene, rather than detecting the interchangeable coding regions. By designing detection primers and probes that specifically bind to these unique flanking sequences, the method achieves event-specific discrimination while ignoring the variable coding portions of different transgenes.
3Adaptability or versatility
If DNA constructs with interchangeable coding regions are used, then manufacturing flexibility is improved, but detection specificity deteriorates
Solution Approach 1:
The invention extracts and targets only the unique flanking DNA sequences adjacent to the inserted transgene, rather than detecting the interchangeable coding regions. By designing detection primers and probes that specifically bind to these unique flanking sequences, the method achieves event-specific discrimination while ignoring the variable coding portions of different transgenes.
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 detection of the DP-915635-4 event in corn plants and progeny, ensuring effective insect resistance and facilitating segregation and quality control of transgenic materials.
Implementation Method 1
a DNA primer set, that when used in a nucleic acid amplification reaction with genomic DNA extracted from corn comprising event DP-915635-4
Implementation Method 2
performing a nucleic acid amplification reaction, thereby producing the amplicon
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-915635-4, and to assays for detecting the presence of event DP-915635-4 in samples and compositions thereof.