title: Coordinate expression and independent subcellular targeting of multiple proteins from a single transgene.
abstract: A variety of conventional methods allow the expression of multiple foreign proteins in plants by transgene stacking or pyramiding. However, most of these approaches have significant drawbacks. We describe a novel alternative, using a single transgene to coordinate expression of multiple proteins that are encoded as a polyprotein capable of dissociating into component proteins on translation. We demonstrate that this polyprotein system is compatible with the need to target proteins to a variety of subcellular locations, either cotranslationally or posttranslationally. It can also be used to coordinate the expression of selectable marker genes and effect genes or to link genes that are difficult to assay to reporter genes that are easily monitored. The unique features of this polyprotein system are based on the novel activity of the 2A peptide of Foot-and-mouth disease virus (FMDV) that acts cotranslationally to effect a dissociation of the polyprotein while allowing translation to continue. This polyprotein system has many applications both as a research tool and for metabolic engineering and protein factory applications of plant biotechnology.
authors.affiliation: Plant Research Unit, School of Life Sciences, University of Dundee, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, Scotland, United Kingdom.
authors.forename: Abdelhak
authors.initials: A
authors.lastname: El Amrani
authors.forename: Abdellah
authors.initials: A
authors.lastname: Barakate
authors.forename: Barak M
authors.initials: BM
authors.lastname: Askari
authors.forename: Xuejun
authors.initials: X
authors.lastname: Li
authors.forename: Alison G
authors.initials: AG
authors.lastname: Roberts
authors.forename: Martin D
authors.initials: MD
authors.lastname: Ryan
authors.forename: Claire
authors.initials: C
authors.lastname: Halpin
mesh_terms: D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D018390:Gene Targeting / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000235:genetics; D019076:Transgenes / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D005819:Genetic Markers; D008565:Membrane Proteins; D010940:Plant Proteins; D014764:Viral Proteins; C057421:virus protein 2A