Modified PIC9 Insecticidal Protein Expression in Maize
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Solution Overview
Problem
Current chemical and biotic insecticides used to control plant pests face limitations such as environmental contamination, drug-resistant insects, and high production costs, while transgenic plants with insect-resistant genes show promise but require effective insecticidal proteins with high expression levels and strong toxicity.
Innovation Solution
Development of a modified PIC9 insecticidal protein and its gene encoding, optimized for high expression and toxicity in plants like maize, using recombinant vectors and genetic engineering to introduce the protein into plants, potentially combined with other insecticidal proteins for enhanced efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chemical insecticides are used to control plant pests, then pest control effectiveness is improved, but environmental contamination and drug-resistant insects occur
Solution Approach 1:
The patent uses transgenic plants that copy the genetic information of Bacillus thuringiensis to produce insecticidal proteins within the plant. This allows the plant itself to produce the protective substance rather than applying external chemical insecticides, thereby maintaining pest control effectiveness while eliminating environmental contamination associated with chemical applications
Solution Approach 2:
The transgenic plant performs self-service by producing its own insecticidal protein through the introduced gene. The plant's cellular machinery synthesizes the protective protein, eliminating the need for external chemical insecticide applications and reducing environmental harm while maintaining effective pest control
2Reliability
If chemical insecticides are applied repeatedly, then pest control is maintained, but production cost increases
Solution Approach 1:
The insecticidal protein is produced in advance within the plant tissue before pest infestation occurs. The protein is stored in crystal structures within the plant, so when pests consume the plant, the toxin is already present and ready to act, eliminating the need for repeated chemical applications and reducing production costs
Solution Approach 2:
The plant serves itself by continuously producing and storing insecticidal proteins within its tissues. This self-providing mechanism eliminates the need for repeated external chemical insecticide applications, thereby maintaining pest control while significantly reducing production costs associated with multiple chemical treatments
3Object-affected harmful factors
If biotic insecticides are used, then environmental friendliness is improved, but they are easily degraded and require repeated application
Solution Approach 1:
The patent modifies the chemical structure and stability parameters of the insecticidal protein by optimizing the gene sequence and protein expression conditions. This results in a more durable protein that resists environmental degradation while maintaining environmental friendliness, extending the duration of action without requiring repeated applications
Solution Approach 2:
The insecticidal protein is produced as a crystalline structure within the plant cell, creating a composite form that combines the biological activity of the protein with the structural stability of crystals. This composite structure protects the protein from environmental degradation while maintaining its insecticidal activity, thereby extending durability without compromising environmental friendliness
4Object-affected harmful factors
If transgenic plants with insect-resistant genes are developed, then chemical insecticide reduction is achieved, but expression level and toxicity need to be optimized
Solution Approach 1:
The patent optimizes the gene parameters including codon usage, promoter strength, and protein processing signals to maximize expression level and toxicity. By carefully adjusting these genetic parameters, the plant produces high concentrations of active insecticidal protein while maintaining safety, thereby achieving both chemical insecticide reduction and reliable protein toxicity
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 modified PIC9 protein demonstrates strong toxicity against Lepidoptera pests, providing high expression levels and stability in maize, reducing the need for chemical insecticides and offering improved protection against insect damage.
Implementation Method 1
Protein N-terminal and C-terminal are digested by alkaline proteinase and the protoxin is converted into active fragments
Implementation Method 2
These active fragments are bounded to receivers on the upper surface of epithelial membrane of insect mesenteron, and inserted into the intestine membrane
Implementation Method 3
which results in cell membrane perforation lesions, destroy of the change of osmotic pressure and the pH balance inside and outside cell membrane
Data Source
AI summary
The invention relates to an insecticidal protein, its gene encoding and the uses thereof. The protein comprises: (a) a protein consisting of an amino acid sequence shown by SEQ ID NO:2; or (b) a protein derived from (a) consisting of an amino acid sequence by substitution, deletion, or addition of one or more amino acid residues of the amino acid sequences in (a), and having insecticidal activity; or (c) a protein generated by the expression of nucleic acid molecules containing a nucleotide sequence of SEQ ID NO:1; or (d) a protein generated by the expression of nucleic acid molecules containing a complementary sequence that hybridized with SEQ ID NO:1 under stringent conditions; or (e) a protein generated by the expression of nucleic acid molecules that contain nucleotide sequences isocoding with the nucleotide sequences in (d). The insecticidal protein of the invention has high expression level and strong toxicity against pests.