Antibodies Detect Mutant HPPD Protein
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Solution Overview
Problem
There is a need for a method to quickly and cost-effectively identify and isolate transgenic plants containing a mutated hydroxyphenylpyruvate dioxygenase (HPPD) protein, which confers tolerance to herbicides, without relying on time-consuming laboratory steps.
Innovation Solution
Development of antibodies that are immunoreactive with the mutated Pseudomonas HPPD polypeptide but not with the soybean wild-type HPPD polypeptide, specifically produced by hybridoma cell lines such as 1C5, 2E6, and 6H11, which can be used to detect and isolate the mutated protein.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional laboratory methods are used to identify transgenic plants containing mutated HPPD protein, then identification accuracy can be achieved, but the process becomes time-consuming and costly
Solution Approach 1:
The invention changes the detection parameter from general protein detection to specific epitope detection. By developing antibodies that recognize unique conformational epitopes created by the mutation (such as the G336W mutation in Pseudomonas HPPD), the method achieves high identification accuracy while enabling rapid detection without time-consuming laboratory procedures. The antibody-based detection transforms the identification process into a straightforward immunological assay.
2Manufacturing precision
If traditional laboratory procedures are used for isolating mutated HPPD protein, then purification precision can be achieved, but the complexity of the process increases
Solution Approach 1:
The invention introduces antibodies as intermediary agents to simplify the purification process. These antibodies specifically bind to the mutated HPPD protein, serving as mediators that enable selective isolation of the target protein from plant extracts. This antibody-based approach replaces complex multi-step purification procedures with a simpler affinity-based isolation method, reducing process complexity while maintaining high purification precision.
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
These antibodies enable rapid identification and isolation of the mutated HPPD protein, facilitating the selection and breeding of transgenic plants with improved herbicide tolerance, reducing the need for extensive field tests and laboratory procedures.
Implementation Method 1
an antibody immunoreactive with a mutated Pseudomonas HPPD polypeptide but not with a soybean wild-type HPPD polypeptide
Data Source
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AI summary
Antibodies immunoreactive to mutant Pseudomonas HPPD are provided, and in an embodiment the mutant HPPD is one in which the wild-type HPPD is substituted at residue 336 with tryptophan for glycine. Also provided are hybridomas producing the antibodies, as well as methods of making and using the antibodies.