Schizochytrium Recombinant Vector for Stable High Gene Expression
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Solution Overview
Problem
Existing technologies face challenges in achieving high and stable expression of introduced genes in Schizochytrium microalgae, with variations in gene expression levels due to random insertion and lack of specific insertion sites, hindering phenotypic changes analysis.
Innovation Solution
A DNA construct is designed with 5' and 3' homology arms to target specific high-expression regions in the Schizochytrium genome, enabling precise gene insertion through homologous recombination, ensuring consistent and high-level gene expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If gene introduction occurs via random insertion into arbitrary sites throughout the genome, then gene introduction is simple and easy to perform, but the expression level of the introduced gene varies among individual transformants
Solution Approach 1:
The invention changes the parameter of insertion site selection from random to specific high-expression regions. By targeting genes to specific genomic loci with known high expression characteristics, the system achieves both efficient gene introduction and consistent high-level expression across transformants, resolving the contradiction between ease of manufacture and manufacturing precision.
Solution Approach 2:
The invention introduces a mediator system consisting of specific genomic target sites with known high-expression properties. These target sites act as intermediaries that receive introduced genes and ensure their consistent high-level expression, thereby maintaining both ease of gene introduction and expression consistency.
2Ease of manufacture
If gene expression level is insufficient, then gene introduction can be performed, but phenotypic changes attributable to the introduced gene cannot be clearly confirmed
Solution Approach 1:
The invention changes the expression level parameter from insufficient to high-level expression by targeting specific genomic regions. This ensures that introduced genes are expressed at sufficiently high levels to produce detectable phenotypic changes, thereby improving measurement precision while maintaining ease of gene introduction.
3Adaptability or versatility
If gene insertion sites are not specific, then gene introduction is flexible, but variation in gene expression among transformants increases
Solution Approach 1:
The invention changes the insertion site parameter from non-specific to specific high-expression regions. This provides a balance between adaptability (multiple available target sites) and stability (consistent expression at each target), resolving the contradiction between insertion flexibility and expression stability.
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 DNA construct achieves stable and high-level expression of target genes in Schizochytrium, allowing for accurate phenotypic changes analysis and efficient production of target proteins or products.
Implementation Method 1
A DNA construct is designed with 5' and 3' homology arms to target specific high-expression regions in the Schizochytrium genome, enabling precise gene insertion through homologous recombination
Data Source
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AI summary
The present application relates to a DNA construct for high expression of a target gene in a microorganism of the genus Schizochytrium, and a method for expressing a target gene in a microorganism of the genus Schizochytrium using same. The DNA construct comprises a 5' homology arm and a 3' homology arm hybridizable with a high-expression site of a microbial genome of the genus Schizochytrium.