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

Fast complementation of split fluorescent protein triggered by DNA hybridization.

Vadim V Demidov1, Nikolay V Dokholyan, Carlos Witte-Hoffmann

  • 1Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.

Proceedings of the National Academy of Sciences of the United States of America
|February 8, 2006
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

Correction: High-resolution DNA size enrichment using a magnetic nano-platform and application in non-invasive prenatal testing.

The Analyst·2022
Same author

Efficient detection and post-surgical monitoring of colon cancer with a multi-marker DNA methylation liquid biopsy.

Proceedings of the National Academy of Sciences of the United States of America·2021
Same author

Artificial intelligence, drug repurposing and peer review.

Nature biotechnology·2020
Same author

ANERGY TO SYNERGY-THE ENERGY FUELING THE RXCOVEA FRAMEWORK.

International journal for multiscale computational engineering·2020
Same author

High-resolution DNA size enrichment using a magnetic nano-platform and application in non-invasive prenatal testing.

The Analyst·2020
Same author

Noninvasive diagnosis of urothelial cancer in urine using DNA hypermethylation signatures-Gender matters.

International journal of cancer·2019

This study introduces a novel split enhanced green fluorescent protein (EGFP) system. Fluorescence is rapidly restored within minutes upon DNA hybridization, overcoming previous hour-long delays.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Split fluorescent proteins act as reporters but require slow chromophore formation for fluorescence restoration.
  • Previous methods for split fluorescent protein reassembly took hours to achieve fluorescence.

Purpose of the Study:

  • To develop a split fluorescent protein system with rapid fluorescence recovery.
  • To demonstrate self-catalytic chromophore formation in a split enhanced green fluorescent protein (EGFP) fragment.
  • To enable fluorescence restoration driven by nucleic acid interactions.

Main Methods:

  • Utilized an N-terminal fragment of enhanced green fluorescent protein (EGFP).
  • Engineered split EGFP fragments linked to complementary oligonucleotides.

Related Experiment Videos

  • Assayed fluorescence restoration kinetics upon mixing fragments.

    • Main Results:

    • Demonstrated self-catalytic fluorogenic chromophore formation within an isolated EGFP fragment.
    • Achieved rapid fluorescence restoration (within minutes) upon DNA hybridization.
    • Showcased the system's responsiveness to complementary oligonucleotide interactions.

    Conclusions:

    • The developed split EGFP system allows for fast, DNA-driven fluorescence restoration.
    • This rapid response system can be applied to detect interaction kinetics in vitro and in living cells.
    • Offers a significant improvement over existing split fluorescent protein technologies for real-time monitoring.