title: High expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants.
abstract: Chloroplast transformation has an extraordinary potential for antigen production in plants because of the capacity to accumulate high levels of recombinant proteins and increased biosafety due to maternal plastid inheritance in most crops. In this article, we evaluate tobacco chloroplasts transformation for the production of a highly immunogenic epitope containing amino acid residues 135-160 of the structural protein VP1 of the foot and mouth disease virus (FMDV). To increase the accumulation levels, the peptide was expressed as a fusion protein with the beta-glucuronidase reporter gene (uidA). The recombinant protein represented the 51% of the total soluble proteins in mature leaves, a level higher than those of the Rubisco large subunit, the most abundant protein in the leaf of a wild-type plant. Despite this high accumulation of heterologous protein, the transplastomic plants and wild-type tobacco were phenotypically indistinguishable. The FMDV epitope expressed in transplastomic plants was immunogenic in mice. These results show that transplastomic tobacco express efficiently the recombinant protein, and we conclude that this technology allows the production of large quantities of immunogenic proteins.
authors.affiliation: Laboratorio de Virología y Biotecnología Vegetal, INGEBI-UBA/CONICET, Ciudad Autónoma de Buenos Aires, Vuelta de Obligado 2490, C1428ADN, Buenos Aires, Argentina.
authors.forename: Ezequiel Matías
authors.initials: EM
authors.lastname: Lentz
authors.forename: María Eugenia
authors.initials: ME
authors.lastname: Segretin
authors.forename: Mauro Miguel
authors.initials: MM
authors.lastname: Morgenfeld
authors.forename: Sonia Alejandra
authors.initials: SA
authors.lastname: Wirth
authors.forename: María José
authors.initials: MJ
authors.lastname: Dus Santos
authors.forename: Marina Valeria
authors.initials: MV
authors.lastname: Mozgovoj
authors.forename: Andrés
authors.initials: A
authors.lastname: Wigdorovitz
authors.forename: Fernando Félix
authors.initials: FF
authors.lastname: Bravo-Almonacid
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry; D002736:Chloroplasts / Q000235:genetics* / Q000821:virology*; D000939:Epitopes / Q000276:immunology*; D005536:Foot-and-Mouth Disease / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D005966:Glucuronidase / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D014026:Nicotiana / Q000235:genetics* / Q000821:virology*; D014170:Transformation, Genetic
chemical_list: D036022:Capsid Proteins; D000939:Epitopes; D010940:Plant Proteins; C442501:VP1 protein, Foot-and-mouth disease virus; D005966:Glucuronidase