Codon-Optimized EPSPS Sequence for Broad Plant Expression
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Solution Overview
Problem
Current glyphosate-resistant transgenic plants face limitations in efficiently expressing heterologous genes across both dicotyledonous and monocotyledonous plant cells, with existing EPSPS sequences not optimized for broad plant expression, leading to suboptimal tolerance to glyphosate and related herbicides.
Innovation Solution
A codon-optimized modified EPSPS sequence is designed, featuring mutations at residues 102 and 106, optimized for expression in both dicotyledonous and monocotyledonous plants, particularly soybeans, to enhance glyphosate resistance by improving codon usage and reducing deleterious sequences and restriction sites, thereby facilitating efficient transcription and translation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing EPSPS sequences are used in transgenic plants, then glyphosate resistance is achieved, but expression efficiency across both dicotyledonous and monocotyledonous plants is limited
Solution Approach 1:
The patent applies parameter changes by optimizing the EPSPS gene sequence at the codon level to match the preferred codon usage patterns of both dicotyledonous and monocotyledonous plants. This involves modifying nucleotide sequences at specific positions (residues 102 and 106) while maintaining the protein's glyphosate resistance function, thereby achieving broad expression efficiency without compromising resistance reliability
2Productivity
If wild-type EPSPS gene is introduced into plants, then basic glyphosate resistance is achieved, but expression is suboptimal due to unoptimized codon usage
Solution Approach 1:
The patent optimizes the EPSPS gene by changing codon parameters to align with host plant preferences. Specifically, residues 102 and 106 are modified to enhance protein stability and expression. This codon optimization increases transcription and translation efficiency, leading to higher protein production levels while maintaining ease of manufacturing the transgenic plants
3Reliability
If mutations are introduced at residues 102 and 106, then glyphosate resistance is enhanced, but the sequence becomes less compatible with wild-type plants
Solution Approach 1:
The patent applies local quality by introducing mutations only at specific positions (residues 102 and 106) of the EPSPS protein where they confer glyphosate resistance, while leaving the rest of the protein sequence intact and compatible with wild-type plants. This localized modification approach enhances resistance reliability without compromising overall adaptability to existing plant genetics
Data Source
AI summary
A nucleic acid molecule encoding a 5-enolpyruvyl-3-phosphoshikimic acid synthase (EPSPS) that provides resistance to glyphosate is provided, that has been optimized for expression in both monocotyledonous and dicotyledonous plants and preferably in soybeans. Methods of use are also provided.