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 Experiment Videos

Engineering a temperature sensitive tobacco etch virus protease.

J Wong1, X Chen1, K Truong1,2

  • 1Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario, Canada M5S 3G9.

Protein Engineering, Design & Selection : PEDS
|October 18, 2017
PubMed
Summary

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

Association of Neisseria cinerea with ocular infections in paediatric patients.

The Journal of infection·1998
Same author

Transcription from the thyroid hormone-dependent promoter of the Xenopus laevis thyroid hormone receptor betaA gene requires a novel upstream element and the initiator, but not a TATA Box.

The Journal of biological chemistry·1998
Same author

Unique organization and involvement of GAGA factors in transcriptional regulation of the Xenopus stromelysin-3 gene.

Nucleic acids research·1998
Same author

Inhibition of cellular proliferation by the Wilms tumor suppressor WT1 requires association with the inducible chaperone Hsp70.

Genes & development·1998
Same author

The value of neck drain in esophageal surgery: a randomized trial.

Diseases of the esophagus : official journal of the International Society for Diseases of the Esophagus·1998
Same author

Unusual features of the Drosophila melanogaster telomere transposable element HeT-A are conserved in Drosophila yakuba telomere elements.

Proceedings of the National Academy of Sciences of the United States of America·1998

Engineered a temperature-sensitive tobacco etch virus protease (tsTEVp) for controlled gene function. This tsTEVp shows efficient activity at 30°C and inactivity at 37°C, enabling precise biological responses.

Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Tobacco etch virus protease (TEVp) is known for its high specificity and efficiency in cleaving target substrates.
  • Controlling enzyme activity is crucial for applications in synthetic biology and genetic engineering.
  • Temperature induction is a common method for modulating gene function due to its rapid response and broad applicability.

Purpose of the Study:

  • To engineer a temperature-sensitive variant of TEVp (tsTEVp) for conditional control of its proteolytic activity.
  • To demonstrate the utility of tsTEVp in generating temperature-inducible biological responses.
  • To explore the potential applications of tsTEVp in engineered synthetic biological systems.

Main Methods:

  • N-terminal truncations were introduced to the TEVp sequence to create a temperature-sensitive variant.
Keywords:
TEV proteasebiosensorlive cell imagingtemperature sensitive

Related Experiment Videos

  • The activity of the engineered tsTEVp was assessed at different temperatures (30°C and 37°C).
  • tsTEVp was utilized to induce biological responses including protein translocation, protein degradation, and Ca2+-mediated cellular blebbing.

    • Main Results:

    • The engineered tsTEVp exhibited efficient proteolysis within 4 hours of induction at 30°C.
    • The tsTEVp remained largely inactive at 37°C, demonstrating temperature-dependent control.
    • Successful induction of protein translocation, degradation, and cellular blebbing was achieved using tsTEVp.

    Conclusions:

    • A functional temperature-sensitive TEVp (tsTEVp) was successfully engineered using N-terminal truncations.
    • tsTEVp provides a reliable tool for precise, temperature-controlled manipulation of biological processes.
    • The developed tsTEVp system holds promise for applications in advanced synthetic biology and genetic engineering.