title: Antigen production using heterologous expression of dengue virus-2 non-structural protein 1 (NS1) in Nicotiana tabacum (Havana) for immunodiagnostic purposes.
abstract: Expression of dengue-2 virus NS1 protein in Nicotiana tabacum plants for development of dengue immunodiagnostic kits. Dengue is one of the most important diseases caused by arboviruses in the world. A significant increase in its geographical distribution has been noticed over the last 20 years, with continuous transmission of several serotypes and emergence of the hemorrhagic fever in areas where the disease was previously not prevalent. Although the methodological processes for dengue diagnosis are in deep development and improvement, a limitation for the realization of dengue diagnostic tests is the difficulty of large-scale production of the antigen to be used in diagnostic tests. Due to this demand, the purpose of this study was to obtain the non-structural protein 1 (NS1) from dengue-2 serotype by heterologous expression in Nicotiana tabacum (Havana). After confirmation of the NS1 protein gene integration in the plant genome, the heterologous protein was characterized using SDS-PAGE and immunoblotting. In an immunoenzymatic test, the recombinant NS1 protein presents an antigen potential for development of dengue immunodiagnostic kits.
authors.affiliation: Laboratory of Molecular Immunovirology, Department of General Biology, Federal University of Viçosa, Av. PH Rolfs, s/n Campus Universitário, Viçosa, MG, CEP 36570-000, Brazil.
authors.forename: Marilane O F
authors.initials: MO
authors.lastname: Amaro
authors.forename: Mariana F
authors.initials: MF
authors.lastname: Xisto
authors.forename: Ana Carolina F
authors.initials: AC
authors.lastname: Dias
authors.forename: Alice F
authors.initials: AF
authors.lastname: Versiani
authors.forename: Silvia A
authors.initials: SA
authors.lastname: Cardoso
authors.forename: Wagner C
authors.initials: WC
authors.lastname: Otoni
authors.forename: Cynthia C
authors.initials: CC
authors.lastname: da Silva
authors.forename: Sérgio O
authors.initials: SO
authors.lastname: De Paula
mesh_terms: D000330:Aedes / Q000821:virology; D016960:Agrobacterium tumefaciens / Q000235:genetics; D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D016022:Case-Control Studies; D003715:Dengue / Q000175:diagnosis*; D003716:Dengue Virus / Q000276:immunology / Q000472:pathogenicity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism
chemical_list: D000956:Antigens, Viral; C079340:NS1 protein, Dengue virus type 2; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins