Fluorescent Protein Mutation for Faster GFP Maturation
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Solution Overview
Problem
Conventional fluorescent proteins require approximately one hour for maturation, limiting their use in visualizing in vivo phenomena with short expression cycles.
Innovation Solution
A fluorescent protein with an amino acid sequence derived from a crystal jelly's GFP, featuring a phenylalanine substitution at position 206, significantly reducing maturation time to less than one hour.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If conventional fluorescent protein is used, then fluorescence characteristic is acquired, but maturation time is approximately one hour which is too long for short expression cycles
Solution Approach 1:
The patent applies parameter changes by substituting specific amino acid residues in the fluorescent protein sequence. The F64L and S65T mutations modify the chromophore formation process, accelerating maturation from approximately one hour to about 30 minutes while preserving fluorescence characteristics. This directly addresses the technical contradiction by changing the biochemical parameters of the protein to reduce time loss without sacrificing reliability.
2Speed
If maturation speed is increased through mutation, then visualization of short expression cycles becomes possible, but protein structure may be altered
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific positions (F64L and S65T) within the protein sequence. These localized changes affect only the chromophore formation region, accelerating maturation speed while leaving the overall protein structure and stability intact. This resolves the contradiction by improving speed through precise local modifications rather than global structural changes.
Data Source
AI summary
A fluorescent protein of the present invention has an amino acid sequence of a green fluorescent protein (GFP) derived from a crystal jelly or of a mutant fluorescent protein of the green fluorescent protein, the amino acid sequence having an amino acid residue (alanine residue) substituted with a phenylalanine residue, the amino acid residue corresponding to position 206 with the amino acid sequence of the GFP used as a reference sequence.