Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Plasmids01:28

Plasmids

83
Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
83
Coordination of Gene Expression Processes in Bacteria01:29

Coordination of Gene Expression Processes in Bacteria

126
The DNA replication, transcription, and translation processes are intricately coupled in bacteria, allowing efficient gene expression and rapid protein synthesis. While this physical and functional coordination is advantageous, it introduces challenges that bacteria overcome through specific regulatory mechanisms.Coupling of Replication, Transcription, and TranslationThe coupling of replication, transcription, and translation is a hallmark of bacterial gene expression. As the replisome unwinds...
126
Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

108
The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
108
Stringent Response in E. coli01:23

Stringent Response in E. coli

40
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...
40

You might also read

Related Articles

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

Sort by
Same author

CRISPR-Associated Transposase for Targeted Mutagenesis in Diverse Proteobacteria.

ACS synthetic biology·2023
Same author

Efficient and iterative retron-mediated <i>in vivo</i> recombineering in <i>Escherichia</i> <i>coli</i>.

Synthetic biology (Oxford, England)·2022
Same author

GuideMaker: Software to design CRISPR-Cas guide RNA pools in non-model genomes.

GigaScience·2022
Same author

Draft Genome Sequences of Actinobacterial and Betaproteobacterial Strains Isolated from the Stratosphere.

Microbiology resource announcements·2021
Same author

A plasmid toolbox for controlled gene expression across the Proteobacteria.

Nucleic acids research·2021
Same journal

Cadmium toxicity to the human gut microbiome varies depending on composition.

Applied and environmental microbiology·2026
Same journal

Structural and functional characterization of dihydrodiol dehydrogenase PahB recognizing high-molecular-weight PAH substrates.

Applied and environmental microbiology·2026
Same journal

<i>Salmonella</i>-derived OMVs as a vaccine platform for <i>Riemerella anatipestifer</i> outer membrane proteins to prevent infection in ducks.

Applied and environmental microbiology·2026
Same journal

Characterization of chemotaxis in soybean symbiont <i>Bradyrhizobium diazoefficiens</i>.

Applied and environmental microbiology·2026
Same journal

Degradation pathway of (R)-1,3-butanediol in <i>Pseudomonas putida</i> KT2440 and development of its biosensor.

Applied and environmental microbiology·2026
Same journal

A single amino acid in the viral capsid makes a major contribution to the interaction between human norovirus GII.6 and the bacterial Psl exopolysaccharide.

Applied and environmental microbiology·2026
See all related articles

Related Experiment Video

Updated: Aug 22, 2025

The Multifaceted Benefits of Protein Co-expression in Escherichia coli
12:48

The Multifaceted Benefits of Protein Co-expression in Escherichia coli

Published on: February 5, 2015

12.1K

Plasmids for Controlled and Tunable High-Level Expression in E. coli.

Layla A Schuster1, Christopher R Reisch1

  • 1Department of Microbiology and Cell Science, University of Florida, Gainesville, Florida, USA.

Applied and Environmental Microbiology
|November 7, 2022
PubMed
Summary
This summary is machine-generated.

This study introduces new inducible gene expression systems that outperform the T7 system for high protein production in bacteria. These novel systems offer better control and higher yields without requiring special strains.

Keywords:
T7 systeminducible expressionplasmid toolboxprotein overexpression

More Related Videos

Controllable Ion Channel Expression through Inducible Transient Transfection
10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

9.5K
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

33.0K

Related Experiment Videos

Last Updated: Aug 22, 2025

The Multifaceted Benefits of Protein Co-expression in Escherichia coli
12:48

The Multifaceted Benefits of Protein Co-expression in Escherichia coli

Published on: February 5, 2015

12.1K
Controllable Ion Channel Expression through Inducible Transient Transfection
10:00

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

9.5K
Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli
08:46

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Published on: January 6, 2015

33.0K

Area of Science:

  • Microbiology
  • Molecular Biology
  • Synthetic Biology

Background:

  • Controlled gene expression is vital for bacterial engineering.
  • The T7 expression system is popular for high protein overexpression but suffers from host toxicity and leaky expression.
  • Existing solutions to T7 limitations often complicate the system or restrict host compatibility.

Purpose of the Study:

  • To challenge the T7 system's dominance for high protein titers.
  • To evaluate alternative inducible systems for improved protein expression and transcriptional control.
  • To demonstrate the utility of a previously developed toolbox of inducible systems.

Main Methods:

  • Comparison of four inducible systems from a plasmid toolbox against the T7 expression system.
  • Assessment of protein output and fold change over basal expression in rich and minimal media.
  • Evaluation of system performance in an engineered metabolic pathway for lycopene production.

Main Results:

  • The developed plasmids consistently yielded higher protein outputs than the T7 system.
  • The alternative systems demonstrated greater fold changes over basal expression compared to T7.
  • The new systems outperformed T7 in lycopene production using an engineered metabolic pathway.

Conclusions:

  • Inducible promoters on high-copy plasmids can rival T7 RNAP expression levels with superior transcriptional control.
  • The plasmid-based, modular systems offer advantages in predictability, stability, and host flexibility over the T7 system.
  • These alternative expression systems expand options for high-level protein expression across diverse bacterial strains.