ArticleAMB Express2018
Rapid monitoring of the target protein expression with a fluorescent signal based on a dicistronic construct in Escherichia coli.
Article in AMB Express, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
18 citing papers in PubMed, 35 citations in OpenAlex.
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- Production and Purification of Cysteine-Rich Leptospiral Virulence-Modifying Proteins with or Without mCherry Fusion.The protein journal · 2023Article
- Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors and tumor-homing bacteria.Nature biotechnology · 2023Article
- A Microfluidic Dual-Aptamer Sandwich Assay for Rapid and Cost-Effective Detection of Recombinant Proteins.Microchemical journal : devoted to the application of microtechniques in all branches of science · 2023Article
- Indirect optimization of staphylokinase expression level in dicistronic auto-inducible system.AMB Express · 2022Article
- Differential Detection of Bioavailable Mercury and Cadmium Based on a Robust Dual-Sensing Bacterial Biosensor.Frontiers in microbiology · 2022Article
- Suppressor Mutants: History and Today's Applications.EcoSal Plus · 2021Review
- A simple and sensitive detection of the binding ligands by using the receptor aggregation and NMR spectroscopy: a test case of the maltose binding protein.Journal of biomolecular NMR · 2021Article
- Indigoidine biosynthesis triggered by the heavy metal-responsive transcription regulator: a visual whole-cell biosensor.Applied microbiology and biotechnology · 2021Article
- Development of a bioavailable Hg(II) sensing system based on MerR-regulated visual pigment biosynthesis.Scientific reports · 2021Article
- Detection of Bioavailable Cadmium by Double-Color Fluorescence Based on a Dual-Sensing Bioreporter System.Frontiers in microbiology · 2021Article
- Versatile artificial mer operons in Escherichia coli towards whole cell biosensing and adsorption of mercury.PloS one · 2021Article
- Novel perspective on a conventional technique: Impact of ultra-low temperature on bacterial viability and protein extraction.PloS one · 2021Article
- Development of Cadmium Multiple-Signal Biosensing and Bioadsorption Systems Based on ArtificialFrontiers in bioengineering and biotechnology · 2021Article
- A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease.AMB Express · 2020Article
- Construction of a RFP-lacZα bicistronic reporter system and its application in lead biosensing.PloS one · 2020Article
- Improved Monitoring of Low-Level Transcription inFrontiers in microbiology · 2019Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Real-time quantification of recombinant proteins is important in studies on fermentation engineering, cell engineering, etc. Measurement of the expression level of heterologous proteins in bacterial fermentation broth has traditionally relied on time-consuming and labor-intensive procedures, such as polyacrylamide gel electrophoresis, immunoblot analysis, and biological activity assays. We describe a simple, fast, and high sensitive assay for detecting heterologous proteins production in bacteria either at the overall level (fluorescence spectrophotometry) or at the individual level (fluorescence microscopic image) in this study. Based on a dicistronic model, the translation of target gene in the upstream open reading frame (ORF) was coupled with the synthesis of the mCherry reporter in the downstream ORF in E. coli cells, and subsequently this demonstrated a positive correlation between the expression of target gene and mCherry. Although a time lag exists between the expression of target protein and mCherry reporter, the method described here allows facile monitoring of dynamic changes in target protein expression, relying on indirect determination of the fluorescence intensity of mCherry during fermentation in real-time models. Additionally, the performance of a single bacterial cell factory could be checked under the fluorescence microscope field.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.