Maize Event DP-0041143 Detection via Flanking Sequence Segmentation
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Solution Overview
Problem
Current methods for detecting specific transgenic events in crops, such as corn event DP-004114-3, are not discriminative enough, particularly when using DNA constructs like those from Bacillus thuringiensis, as they often rely on genetic elements like promoters or terminators, which can be interchangeable, making it difficult to distinguish between different events.
Innovation Solution
A DNA construct comprising the cry1F, cry34Ab1, cry35Ab1, and pat gene cassettes, expressed under specific promoters and terminators, is introduced into maize plants to confer insect resistance, along with methods using specific primers or probes that recognize unique flanking sequences of DP-004114-3 for accurate detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If DNA constructs with interchangeable genetic elements (promoters, terminators) are used for transgenic event detection, then detection simplicity is improved, but detection precision deteriorates because different events cannot be distinguished
Solution Approach 1:
The detection method is segmented into two distinct parts: (1) detection of common genetic elements (promoters, terminators, selectable markers) that are interchangeable across events, and (2) detection of unique flanking sequences that are specific to each transgenic event. This segmentation allows the method to maintain simplicity for common elements while achieving precision through event-specific flanking sequence detection, thereby resolving the contradiction between ease of operation and measurement precision.
2Reliability
If screening of large number of events is performed to identify optimal transgene expression, then expression reliability is improved, but time consumption and resource usage increase
Solution Approach 1:
The patent employs preliminary action by using molecular markers and flanking sequence detection to identify and select specific transgenic events with desired expression characteristics before large-scale screening is performed. This preliminary identification through PCR-based detection of event-specific flanking sequences allows researchers to narrow down the candidate events, thereby maintaining expression reliability while significantly reducing the time and resources required for comprehensive screening of hundreds or thousands of events.
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
The approach enables effective insect resistance in maize plants and provides a discriminative method for identifying the DP-004114-3 event, ensuring reliable gene expression and compliance with regulatory requirements.
Implementation Method 1
A DNA construct comprising the cry1F, cry34Ab1, cry35Ab1, and pat gene cassettes, expressed under specific promoters and terminators, is introduced into maize plants to confer insect resistance
Implementation Method 2
methods using specific primers or probes that recognize unique flanking sequences of DP-004114-3 for accurate detection
Data Source
AI summary
The invention provides DNA compositions that relate to transgenic insect resistant maize plants. Also provided are assays for detecting the presence of the maize DP-004114-3 event based on the DNA sequence of the recombinant construct inserted into the maize genome and the DNA sequences flanking the insertion site. Kits and conditions useful in conducting the assays are provided.