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

You might also read

Related Articles

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

Sort by
Same author

<i>Clostridioides difficile</i> Stimulates <i>CCL20</i> Expression in Human Colonoid Monolayers in a Transwell-Based Coculture System That Supports Its Anaerobic Growth.

International journal of microbiology·2026
Same author

Naturally Occurring CodY Variants Alter Ligand Binding, DNA Target Affinity, and Virulence in Clostridioides difficile.

Molecular microbiology·2026
Same author

<i>Clostridioides difficile</i> stimulates <i>CCL20</i> expression in human colonoid monolayers in a transwell-based co-culture system that supports its anaerobic growth.

bioRxiv : the preprint server for biology·2026
Same author

Overexpression of the CYP81Q32-like gene in wheat (Triticum aestivum) reduces sensitivity to triketone herbicides.

Pesticide biochemistry and physiology·2025
Same author

Regulatory networks: Linking toxin production and sporulation in Clostridioides difficile.

Anaerobe·2024
Same author

Erratum for Hasan et al., "Role of glycogen metabolism in <i>Clostridioides difficile</i> virulence".

mSphere·2024

Related Experiment Video

Updated: Dec 21, 2025

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology
16:11

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Published on: September 16, 2013

65.7K

A simplified method for producing laboratory grade recombinant TEV protease from E. coli.

Jordan Brungardt1, Revathi Govind2, Harold N Trick1

  • 1Department of Plant Pathology, Kansas State University, Manhattan, KS 66502, USA.

Protein Expression and Purification
|May 11, 2020
PubMed
Summary

We developed a faster method to produce tobacco etch virus (TEV) protease in E. coli. This expedited process uses freeze-thaw lysis and affinity chromatography for high-purity protease essential for fusion protein cleavage.

Keywords:
Fusion protein cleavageRecombinant protein isolationTEV proteasepDZ2087

More Related Videos

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
08:09

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Published on: March 24, 2017

9.8K
Author Spotlight: The Production of Recombinant Proteins
05:19

Author Spotlight: The Production of Recombinant Proteins

Published on: June 30, 2023

9.4K

Related Experiment Videos

Last Updated: Dec 21, 2025

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology
16:11

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Published on: September 16, 2013

65.7K
Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
08:09

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

Published on: March 24, 2017

9.8K
Author Spotlight: The Production of Recombinant Proteins
05:19

Author Spotlight: The Production of Recombinant Proteins

Published on: June 30, 2023

9.4K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Chemistry

Background:

  • Tobacco etch virus (TEV) protease is crucial for cleaving fusion proteins due to its specific sequence recognition.
  • Traditional methods for isolating recombinant proteins from E. coli involve harsh cell lysis techniques like sonication or French press.

Purpose of the Study:

  • To develop an expedited and efficient laboratory-scale method for producing high-purity TEV protease in E. coli.
  • To optimize the purification process using immobilized metal affinity chromatography (IMAC).

Main Methods:

  • TEV protease was expressed in BL21 (DE3) E. coli cells using the pDZ2087 plasmid.
  • A freeze-thaw lysis method was employed for cell disruption, replacing conventional harsh methods.
  • Purification was achieved using immobilized metal affinity chromatography.

Main Results:

  • The freeze-thaw method combined with IMAC successfully produced laboratory-grade TEV protease.
  • The purified TEV protease demonstrated high efficiency, cleaving maltose-binding protein fusions completely.
  • Optimal cleavage was observed at a fusion-to-protease molar ratio of 50:1.

Conclusions:

  • The developed method offers a faster and gentler alternative for producing active TEV protease.
  • This expedited protocol simplifies recombinant protein purification workflows in research settings.
  • The high efficiency of the produced TEV protease ensures effective fusion protein cleavage.