Abstract
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 β-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 CAT2A-GUS 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.
| Original language | English |
|---|---|
| Pages (from-to) | 928-933 |
| Number of pages | 6 |
| Journal | EMBO Journal |
| Volume | 13 |
| Issue number | 4 |
| Publication status | Published - 15 Feb 1994 |
Keywords
- Autoproteolytic
- Cotranslational cleavage
- Foot-and-mouth disease virus
- Polyprotein