What question did this study set out to answer?

To establish an Escherichia coli dual-plasmid expression platform that prevents leaky expression, resolves codon bias, and promotes protein folding without compromising cloning plasmid yield.

September 17, 2026Applied Microbiology and BiotechnologyOpen Access

A dual-plasmid system for efficient soluble protein expression in Escherichia coli

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Authors

YCYin ChenZhejiang Ocean University JLJialin LuoGuilin Medical University HLHan LiKunming Medical University

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Implication

Experimental study demonstrates a dual-plasmid system controls leaky expression and enhances protein solubility in Escherichia coli, indicating a versatile tool for recombinant protein production.

Key Points

To establish an Escherichia coli dual-plasmid expression platform that prevents leaky expression, resolves codon bias, and promotes protein folding without compromising cloning plasmid yield.
Engineered an expression plasmid (pSE2) paired with an auxiliary plasmid (pRARE2a-GKJE) encoding Rop protein, LacI repressor, rare tRNAs, and molecular chaperones.
Assessed plasmid copy number regulation, leaky transcription, and soluble expression efficiency across human-codon-optimized genes, alongside direct eukaryotic functional validation.
pSE2 sustained high copy numbers alone for DNA preparation but underwent Rop-mediated copy number suppression and LacI-mediated repression when co-expressed with pRARE2a-GKJE.
Achieved high soluble expression yields across challenging targets: up to 97% for TurboID, 99% for Spy Cas9, and 53% for PE6d.
Enabled direct eukaryotic functional validation using the pSE2 construct without requiring re-cloning.

Cite This Study

Chen et al. (2026) studied this question.

synapsesocial.com/papers/6aabb7755f706d05830e6841 https://doi.org/10.1007/s00253-026-14039-x

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Implication

Experimental study demonstrates a dual-plasmid system controls leaky expression and enhances protein solubility in Escherichia coli, indicating a versatile tool for recombinant protein production.

A dual-plasmid system for efficient soluble protein expression in Escherichia coli

Discussion

Member takes

Implication

Key Points

Cite This Study

A dual-plasmid system for efficient soluble protein expression in Escherichia coli

Discussion

Member takes

Implication

Key Points

Cite This Study

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