Soybean Event Detection via Flanking Sequence Primers
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Solution Overview
Problem
Current methods for detecting specific transgenic soybean events, such as Soybean Event pDAB9582.816.15.1, are not discriminative enough, particularly when multiple events are produced using similar DNA constructs, making it difficult to identify and verify the presence of specific transgenes in plants and their progeny, which is crucial for regulatory compliance and breeding purposes.
Innovation Solution
A method involving specific PCR primers that bind to unique flanking and insert sequences surrounding the transgene insertion site in the soybean genome, allowing for the detection of Soybean Event pDAB9582.816.15.1 by amplifying diagnostic amplicons, and the use of DNA sequences and probes for accurate identification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If general transgene detection methods (PCR or DNA hybridization) are used, then detection capability is achieved, but discrimination between different events is insufficient
Solution Approach 1:
The detection method segments the target DNA into specific regions of interest (flanking sequences and junction sequences) and designs primers/probes that specifically bind to these segmented regions. This segmentation enables the detection system to distinguish between different transgenic events by targeting unique sequence combinations rather than detecting all transgenes generically.
Solution Approach 2:
The invention applies local quality by designing detection reagents with specific binding characteristics for different genomic regions. The primers and probes are tailored to bind to specific flanking sequences and junction sequences that are unique to each transgenic event, providing localized discrimination capability rather than uniform detection across all events.
2Adaptability or versatility
If event-specific detection is implemented, then discrimination between events is improved, but complexity of detection method increases
Solution Approach 1:
The detection method achieves universality by using a standardized PCR-based platform that can detect multiple different transgenic events. The same basic detection system and equipment are used, but different primer and probe combinations allow the system to universally detect various events without requiring separate complex detection systems for each event.
Solution Approach 2:
The invention simplifies complexity by changing the parameters of the detection reagents (primers and probes) rather than changing the fundamental detection methodology. By modifying the sequence parameters of the primers and probes to match different flanking and junction sequences, the system can detect different events using the same operational framework and equipment.
3Reliability
If screening of large number of events is performed, then optimal expression event is identified, but time and resources are consumed
Solution Approach 1:
The invention applies preliminary action by performing event-specific detection early in the breeding and screening process. The method enables rapid identification of events with desired expression characteristics before committing significant resources to further development, allowing researchers to preliminarily screen and select promising events efficiently.
Solution Approach 2:
The invention replaces time-consuming manual screening and phenotypic analysis with a molecular-based detection system. By substituting mechanical/phenotypic observation with PCR-based molecular detection, the method accelerates the screening process while maintaining reliability in identifying events with optimal transgene expression.
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 Soybean Event pDAB9582.816.15.1 in soybean samples, ensuring compliance with regulations and facilitating breeding programs by providing a reliable method for detecting the presence of specific transgenes, even in progeny plants.
Implementation Method 1
DNA hybridization using nucleic acid probes
Implementation Method 2
the polymerase chain reaction (PCR)
Data Source
AI summary
Soybean Event pDAB9582.816.15.1 comprises gene expression cassettes which contain genes encoding Cry1F, Cry1Ac (synpro), and PAT, affording insect resistance and herbicide tolerance to soybean crops containing the event, and enabling methods for crop protection and protection of stored products. The disclosure provides polynucleotide related event detection methods. The present disclosure relates to a method for detecting a new insect resistant and herbicide tolerant transgenic soybean transformation event, designated Soybean Event pDAB9582.816.15,1. The DNA of soybean plants containing this event includes the junction/flanking sequences described herein that characterize the location of the inserted DNA within the soybean genome. SEQ ID NO: 1 and SEQ ID N0:2 are diagnostic for Soybean Event pDAB9582.816.15.1.