title: Production of interleukin-12 as a self-processing 2A polypeptide.
abstract: Interleukin-12 (IL-12) is a heterodimeric cytokine composed of two disulfide-linked subunits (p40 and p35) encoded by separate genes. We used the apparent autocleavage property of a 2A peptide from the foot-and-mouth disease virus (FMDV) to express bovine (Bo) IL-12 as a self-processing polypeptide (p402Ap35). We demonstrate that 2A will mediate the cleavage of p402Ap35 into two separate subunits in a manner similar to that observed during the processing of the FMDV polypeptide. Furthermore, this 2A polypeptide encoded a functional heterodimer, which elicited activities associated with IL-12 in other species. We propose that this strategy of self-processing polypeptides may be used in many applications where the coordinated and stoichiometric expression of complex proteins is required.
authors.affiliation: Institute for Animal Health, Compton, Nr. Newbury, Berkshire, UK. paul.chaplin@aahl.dah.csiro.au
authors.forename: P J
authors.initials: PJ
authors.lastname: Chaplin
authors.forename: E B
authors.initials: EB
authors.lastname: Camon
authors.forename: B
authors.initials: B
authors.lastname: Villarreal-Ramos
authors.forename: M
authors.initials: M
authors.lastname: Flint
authors.forename: M D
authors.initials: MD
authors.lastname: Ryan
authors.forename: R A
authors.initials: RA
authors.lastname: Collins
mesh_terms: D000818:Animals; D019556:COS Cells; D002417:Cattle; D002478:Cells, Cultured; D005822:Genetic Vectors; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D014764:Viral Proteins; C057421:virus protein 2A; D018664:Interleukin-12