Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Stringent Response in E. coli

Stringent Response in E. coli

Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

iChip

iChip

The cultivation of environmental microorganisms has long been hindered by the inability to replicate complex native conditions in vitro. The isolation chip (iChip) addresses this limitation by facilitating the growth of previously uncultivable microorganisms through in situ incubation. Designed for high-throughput microbial cultivation, the iChip comprises hundreds of microchambers, each capable of housing a single microbial cell. These microchambers are loaded with a mixture of molten agar and...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Assembly and Reactions of Artificial Metalloenzymes in <i>Streptomyces albus</i>.

Journal of the American Chemical Society·2026

Same author

TridentSynth: a webtool for the retrosynthesis of molecules using chimeric type I polyketide synthases and chemoenzymatic pathways.

Nucleic acids research·2026

Same author

Influence of titer, rate, yield, and scale on the cost and life-cycle emissions of biomanufacturing.

Trends in biotechnology·2026

Same author

Multi-layered metabolic remodeling of <i>Pseudomonas putida</i> for efficient conversion of lignocellulosic sugars to the precursors of advanced aviation fuel.

Metabolic engineering communications·2026

Same author

Origin of replication discovery for environmentally isolated <i>Pantoea</i> strain enables expression of heterologous proteins, pathways and products.

iScience·2026

Same author

Evaluating isoprenol production using the IPP-bypass pathway in the oleaginous yeast Rhodosporidium toruloides.

Biotechnology for biofuels and bioproducts·2026

Search research articles

Related Experiment Video

Updated: Apr 27, 2026

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat

Published on: September 20, 2016

Development of a native Escherichia coli induction system for ionic liquid tolerance.

Marijke Frederix¹, Kimmo Hütter¹, Jessica Leu¹

¹Joint BioEnergy Institute, Emeryville, California, United States of America; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, United States of America.

|July 2, 2014

Summary

Researchers identified native E. coli promoters to control expression of an ionic liquid (IL) efflux pump gene. This engineering enhances microbial tolerance to ILs, crucial for biofuel production from biomass.

More Related Videos

Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System

Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System

Published on: January 5, 2024

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli

Published on: August 29, 2025

Related Experiment Videos

Last Updated: Apr 27, 2026

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat

Published on: September 20, 2016

Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System

Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System

Published on: January 5, 2024

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli

Published on: August 29, 2025

Area of Science:

Biotechnology
Synthetic Biology
Microbial Engineering

Background:

Ionic liquids (ILs) are effective for lignocellulose pretreatment in biofuel production.
Residual ILs in biomass hydrolysates inhibit microorganism growth and function.
Heterologous expression of the eilA gene in E. coli can confer tolerance to ILs.

Purpose of the Study:

To identify native E. coli IL-inducible promoters for regulating eilA gene expression.
To develop novel control systems for enhancing microbial tolerance to ILs.
To compare the efficacy of native promoters against an IPTG-inducible system.

Main Methods:

Microarray analysis to identify IL-inducible promoters.
Engineering E. coli strains with eilA under control of candidate promoters (PmarR', PydfO', PydfA').
Comparison with PlacUV5 IPTG-inducible system.
Selected-reaction monitoring mass spectrometry for protein quantification.
Pooled culture competition assays to assess strain fitness.

Main Results:

Native promoters PydfA' and PmarR' effectively rescued E. coli from IL toxicity, comparable to PlacUV5.
Mechanistic model indicated inducible systems reduce gene expression at low IL levels.
PydfA' and PmarR' significantly elevated EilA protein levels at high IL concentrations.
The pPmarR'-eilA strain demonstrated superior fitness and outcompeted other strains at IL concentrations >150 mM.

Conclusions:

Native E. coli promoters, particularly PmarR', can effectively regulate heterologous gene expression for IL tolerance.
These native systems eliminate the need for IPTG induction, simplifying strain development.
The findings facilitate the creation of industrially robust microbial strains for biofuel production.