Expressão do peptídeo vacina SBm7462® anti-carrapato Rhipicephalus microplus em plantas transgênicas (Arabidopsis thaliana)

• Ano de defesa: 2010
• Autor(a) principal: Peconick, Ana Paula
• Orientador(a): Salcedo, Joaquín Hernán Patarroyo
• Banca de defesa: Bechara, Gervásio Henrique , Passos, Flávia Maria Lopes , Portela, Ricardo Wagner Dias , Oliveira, Leandro Licursi de
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Doutorado em Medicina Veterinária
• Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
• País: BR
• Palavras-chave em Português: Vacinação; Proteínas heterólogas; Plantas transgênicas
• Palavras-chave em Inglês: Vaccination; Heterologous proteins; Transgenic plants
• Área do conhecimento CNPq: CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA
• Link de acesso: http://locus.ufv.br/handle/123456789/1433

The demand for parasitic vaccines in animal health is growing. The damage due to infestation of ticks and diseases transmitted by them are in the billions of dollars, and the tick Rhipicephalus microplus parasite of the greater economic impact on livestock in Brazil and in countries with tropical and subtropical climates. Advanced research projects, applying tools of biotechnology are used in some or all stages in the development of vaccines. The production of heterologous proteins is a promising alternative for the manufacture of vaccines. The plants represent one of the most economical for large-scale production of proteins for industrial and pharmaceutical, which means using plants as biofactories. The objective of this work is to produce vaccine antigens homologous synthetic peptide SBm7462® against the tick R. microplus in plants (Arabidopsis thaliana) transformed with synthetic gene. The gene seq2, which has the sequence SBm7462® repeated three times in tandem was inserted into donor vector (pDONR201) and then in binary expression vectors for plants (pk7WG2 and pk7FWG2), using the Gateway® system. By selection in medium with antibiotics and polymerase chain reaction (PCR) were selected as positively transformed plants, the presence of these transgene was confirmed by genetic sequencing. Transgenic plants with improved development presented seq2 gene integrated in its genome. To evaluate the expression of the recombinant peptide were carried out RT-PCR (reverse transcription-polymerase chain reaction), immunohisto chemistry and confocal microscope analysis. The recombinant proteins were expressed rBm7462 the apoplast of epidermal cells of transgenic leaves. Plants with the highest levels of expression were selected for new generations, to immunize BALB/c. This is the first study of the production of subunit vaccine in transgenic plants against an ectoparasite.