Foot-and-mouth disease virus 2A oligopeptide mediated cleavage of an artificial polyprotein

EMBO J. 1994 Feb 15;13(4):928-33. doi: 10.1002/j.1460-2075.1994.tb06337.x.


We describe the construction of a plasmid (pCAT2AGUS) encoding a polyprotein in which a 19 amino acid sequence spanning the 2A region of the foot-and-mouth disease virus (FMDV) polyprotein was inserted between the reporter genes chloramphenicol acetyl transferase (CAT) and beta-glucuronidase (GUS) maintaining a single, long open reading frame. Analysis of translation reactions programmed by this construct showed that the inserted FMDV sequence functioned in a manner similar to that observed in FMDV polyprotein processing: the CAT2AGUS polyprotein underwent a cotranslational, apparently autoproteolytic, cleavage yielding CAT-2A and GUS. Analysis of translation products derived from a series of constructs in which sequences were progressively deleted from the N-terminal region of the FMDV 2A insertion showed that cleavage required a minimum of 13 residues. The FMDV 2A sequence therefore provides the opportunity to engineer either whole proteins or domains such that they are cleaved apart cotranslationally with high efficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Aphthovirus / metabolism*
  • Base Sequence
  • Cells, Cultured
  • Chloramphenicol O-Acetyltransferase / genetics
  • DNA, Viral
  • Glucuronidase / genetics
  • Humans
  • Hydrolysis
  • Molecular Sequence Data
  • Peptides / chemical synthesis
  • Peptides / metabolism*
  • Plasmids
  • Protein Biosynthesis
  • Protein Processing, Post-Translational
  • Rabbits
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*


  • DNA, Viral
  • Peptides
  • Viral Proteins
  • virus protein 2A
  • Chloramphenicol O-Acetyltransferase
  • Glucuronidase