Expressão heteróloga do domínio III da proteína de envolope (E) do vírus dengue-1 e 2 em Pichia pastoris

• Ano de defesa: 2011
• Autor(a) principal: Oliveira, André Silva de
• Orientador(a): Paula, Sérgio Oliveira de
• Banca de defesa: Silva, Cynthia Canedo da , Honda, Eduardo Rezende
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Mestrado em Biologia Celular e Estrutural
• Departamento: Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos
• País: BR
• Palavras-chave em Português: Dengue; Expressão heteróloga; Proteína E
• Palavras-chave em Inglês: Dengue; Heterologous expression; E protein
• Área do conhecimento CNPq: CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
• Link de acesso: http://locus.ufv.br/handle/123456789/2332

Annually, the dengue virus (DENV) are responsible for 100 million infections worldwide. The virus has four distinct serotypes, DENV-1, 2, 3 and 4, and consists of three structural proteins (C, prM and E) and seven non-structural. The envelope protein (E) is responsible for binding the virus to the host cell, consisting of three domains (I, II and III), where stands the domain III (DIII) because it presents "immunoglobulin-like" conformation, and it is considered the most immunostimulatory component. The definitive diagnosis of infection by dengue virus can only be done in the laboratory and depends of the isolation of these viruses, detection of viral antigens or RNA in serum or tissue, or detection of specific antibodies in the serum of patients. This process is time consuming, which hinders the epidemiological and clinical intervention. The rapid and cheap diagnostic kits is the solution for this, but the biggest obstacle to its production has been getting immunostimulating proteins in quantity and quality. In this work, we used a heterologous expression system for the production of domain III of envelope protein (E) of dengue virus 1 and 2. The yeast Pichia pastoris has been adopted for the expression of this protein domain because it presenting a pattern of post-translational modifications similar to that of mammals. In our results, molecular tests confirmed the integration of the DIII protein gene in the yeast genome. Preliminary tests about the expression presented encouraging results, it showing that the heterologous expression in Pichia pastoris is promising as the production of antigens of biotechnological interest.