title: Expression of the rabies virus nucleoprotein in plants at high-levels and evaluation of immune responses in mice.
abstract: Transgenic plants have been employed successfully as a low-cost system for the production of therapeutically valuable proteins including antibodies, antigens and hormones. Here, we report expression of a full-length nucleoprotein gene of rabies virus in transgenic tomato plants. The nucleoprotein was also transiently expressed in Nicotiana benthamiana plants by agroinfiltration. In both cases, the nucleoprotein was expressed at high levels, 1-5% of total soluble protein in tomato and 45% in N. benthamiana. Previously, only epitopes of the nucleoprotein had been expressed in plants. The presence and expression of the transgene was verified by PCR, Southern, northern and western blots. Mice were immunized both intraperitoneally (i.p.) and orally with tomato protein extracts containing the N protein induced the production of antibodies. The antibody titer of mice immunized i.p., was at least four times higher than that of mice immunized orally. These results were reflected in the challenge experiments where i.p.-immunized mice were partially protected against a peripheral virus challenge whereas orally immunized mice were not. This protection was comparable to that obtained in previous experiments employing different expression systems. Work is in progress to express both G and N proteins in transgenic plants and evaluate protection in mice.
authors.affiliation: Departamento de Ingeniería Genética de Plantas, Libramiento Norte Carretera Irapuato-León, Irapuato, 36500, Guanajuato, Mexico.
authors.forename: Irene
authors.initials: I
authors.lastname: Perea Arango
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Loza Rubio
authors.forename: Edith
authors.initials: E
authors.lastname: Rojas Anaya
authors.forename: Teresa
authors.initials: T
authors.lastname: Olivera Flores
authors.forename: Luis
authors.initials: L
authors.lastname: Gonzalez de la Vara
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Gómez Lim
mesh_terms: D000818:Animals; D000917:Antibody Formation; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D007114:Immunization; D018551:Solanum lycopersicum / Q000378:metabolism*; D051379:Mice; D009698:Nucleoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D030821:Plants, Genetically Modified / Q000378:metabolism; D011818:Rabies / Q000276:immunology / Q000517:prevention & control; D011820:Rabies virus / Q000276:immunology / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014026:Nicotiana / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D009698:Nucleoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins