Soybean pEIF1 Promoters for Stable Foreign Gene Expression
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Solution Overview
Problem
Existing soybean promoters, such as the 35S promoter, lead to transgenic silencing and safety issues, and constitutive promoters like Gmubi are often stage-specific, leading to challenges in multiple gene insertion and expression in soybean tissues, particularly in root nodules.
Innovation Solution
Utilization of the soybean reference genes RPS28 and EIF1, along with their promoters pRPS28, pRPS28-I, pEIF1, and pEIF1-I, to drive constitutive or non-tissue-specific expression of foreign genes in soybean, Arabidopsis, and tobacco, using recombinant vectors like pRPS28-GUS-BAR, pRPS28-I-GUS-BAR, pEIF1-GUS-BAR, and pEIF1-I-GUS-BAR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral promoters such as 35S or CMV are used for soybean transformation, then high-level gene expression is achieved, but transgenic silencing and safety problems occur
Solution Approach 1:
The patent extracts and utilizes the endogenous soybean promoter pGmUbi from the soybean genome itself, replacing viral promoters. This extraction of the promoter from the host organism's own genetic material eliminates the foreignness that triggers silencing mechanisms while maintaining high expression levels.
Solution Approach 2:
The patent modifies promoter parameters by using the endogenous pGmUbi promoter which naturally maintains high expression levels in soybean. The promoter sequence and its regulatory elements are optimized to achieve constitutive expression without the instability associated with viral promoters.
2Productivity
If constitutive promoters like Gmubi are used, then high expression levels are achieved, but they are stage-specific and rarely active during nodule formation
Solution Approach 1:
The patent enhances the versatility of the pGmUbi promoter by demonstrating its universal activity across multiple developmental stages and tissue types, including roots, nodules, leaves, and seeds. The promoter functions constitutively throughout plant development, making it applicable for various genetic engineering purposes.
Solution Approach 2:
The patent utilizes the dynamic expression pattern of pGmUbi which adapts to different developmental contexts. The promoter maintains activity during nodule formation and other critical developmental stages, responding to physiological conditions while ensuring consistent expression.
3Adaptability or versatility
If multiple genes are inserted into the soybean genome, then enhanced functionality is achieved, but transgenic silencing occurs
Solution Approach 1:
The patent extracts the promoter from endogenous soybean genes and uses it to drive multiple transgene expressions. This endogenous promoter avoids the silencing mechanisms that are triggered by repeated viral promoter sequences, enabling stable co-expression of multiple foreign genes.
Solution Approach 2:
The patent changes the promoter parameter from viral to endogenous origin, which fundamentally alters the interaction with plant defense and silencing mechanisms. This parameter change allows multiple genes to be expressed stably without triggering transcriptional silencing.
Data Source
AI summary
A method for promoting expression of a foreign gene in a plant includes: constructing a recombinant vector including a full-length soybean gene promoter pEIF1 or an intron-including soybean gene promoter pEIF1-I, and introducing the recombinant vector into the plant; the full-length promoter pEIF1 being represented by SEQ ID NO: 5, and the intron-including promoter pEIF1-I being represented by SEQ ID NO: 6.