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

Targeting of proteins derived from self-processing polyproteins containing multiple signal sequences.

Pablo de Felipe1, Martin D Ryan

  • 1School of Biology, Centre for Biomolecular Sciences, Biomolecular Sciences Building, University of St Andrews, North Haugh, St Andrews KY16 9ST, UK.

Traffic (Copenhagen, Denmark)
|July 21, 2004
PubMed
Summary

The 2A peptide from foot-and-mouth disease virus enables co-expression of multiple proteins from one genetic unit. This system allows proteins to be targeted to specific cellular locations, even when linked, facilitating rapid cell line development.

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

Stereotypical thinking in the historiography of science and religion: creation and reception history of Reijer Hooykaas's <i>Religion and the Rise of Modern Science</i>.

Annals of science·2026
Same author

The Coming of Age of Gene Therapy for the Treatment of Human Diseases: A Regulatory Perspective.

Human gene therapy·2025
Same author

Picornavirus Evolution: Genomes Encoding Multiple 2A<sup>NPGP</sup> Sequences-Biomedical and Biotechnological Utility.

Viruses·2024
Same author

Immunogenicity of a secreted, C-terminally truncated, form of bovine viral diarrhea virus E2 glycoprotein as a potential candidate in subunit vaccine development.

Scientific reports·2023
Same author

Mutagenesis Mapping of RNA Structures within the Foot-and-Mouth Disease Virus Genome Reveals Functional Elements Localized in the Polymerase (3D<sup>pol</sup>)-Encoding Region.

mSphere·2021
Same author

A Transgenic Line That Reports CSF1R Protein Expression Provides a Definitive Marker for the Mouse Mononuclear Phagocyte System.

Journal of immunology (Baltimore, Md. : 1950)·2020

Area of Science:

  • Molecular Biology
  • Protein Expression Systems
  • Cellular Biology

Background:

  • The 2A self-cleaving oligopeptide from foot-and-mouth disease virus facilitates co-expression of multiple proteins from a single open reading frame (ORF).
  • 2A mediates co-translational ribosomal skipping at its C-terminus, creating a peptide backbone discontinuity rather than undergoing proteolysis.

Purpose of the Study:

  • To investigate the utility of the 2A peptide system for targeting discrete protein processing products.
  • To explore the co-translational translocation of polyproteins containing the 2A element and signal sequences into the endoplasmic reticulum.

Main Methods:

  • Construction of self-processing polyproteins with fluorescent proteins flanking the 2A sequence.
  • Permutation of signal sequence types and their locations within the polyprotein.

Related Experiment Videos

  • Analysis of protein translocation into the endoplasmic reticulum for polyproteins with and without signal sequences.

    • Main Results:

    • A polyprotein with an N-terminal signal sequence, followed by 2A and a second protein lacking a signal sequence, resulted in both proteins being translocated into the endoplasmic reticulum.
    • The mammalian ribosome:translocon complex remained intact, allowing the second protein to 'slipstream' through the translocon formed by the first protein.
    • Successful targeting of discrete proteins to the endoplasmic reticulum using a single ORF.

    Conclusions:

    • The 2A polyprotein system offers a novel method for targeting proteins to different subcellular locations via single ORF transfection.
    • Fluorescent reporters aid in visualizing expression levels, and selectable markers facilitate rapid establishment of stable cell lines.