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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2011.tde-10022012-120107
Document
Author
Full name
Luciana Moreira Martins
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Mendonça, Ronaldo Zucatelli (President)
Jerez, José Antonio
Wolff, José Luiz Caldas
Title in Portuguese
Estudo do efeito de uma proteína antiapoptótica obtida da hemolinfa de Lonomia obliqua sobre as mitocôndrias de células Sf-9.
Keywords in Portuguese
Baculoviridae
Lonomia oblíqua
Apoptose
Hemolinfa
Mitocôndrias
Proteínas recombinantes
Abstract in Portuguese
O sistema de expressão de proteínas que utiliza baculovírus como vetor tem sido intensamente utilizado para a produção de proteínas recombinantes, pois ele permite a formação de modificações pós-traducionais e conta com um forte promotor, a poliedrina. Porém, em alguns casos, esse sistema não é tão eficaz, pois a infecção das células pelo vírus induz morte por apoptose, limitando a produção industrial de várias proteínas recombinantes de interesse. Reportamos a ocorrência de apoptose em cultivos de células de insetos e de mamíferos com depleção de nutrientes ou induzidos por agentes químicos e virais, e a suplementação dessas culturas com hemolinfa de Lonomia obliqua é capaz de estender a viabilidade da cultura evitando a morte por apoptose. Como objetivo, verificamos previamente o mecanismo de ação antiapoptótica de um isolado protéico da hemolinfa de L. obliqua. A identificação deste mecanismo poderá auxiliar no desenvolvimento de estratégias para controlar a apoptose nos cultivos permitindo a otimização da produtividade viral e de proteínas recombinantes.
Title in English
Effect of an anti-apoptotic protein obtained from the hemolymph of Lonomia obliqua on the mitochondria of cells Sf-9.
Keywords in English
Baculoviridae
Lonomia obliqua
Apoptosis
Hemolymph
Mitochondria
Recombinant proteins
Abstract in English
The protein expression system using baculovirus as a vector has been intensively used for the production of recombinant proteins because it allows the formation of post-translational modifications and has a strong promoter, polyhedrin. However, in some cases, this expression system is not as effective because the infection of cells by baculovirus induces death by apoptosis, thereby limiting the industrial production of various recombinant proteins of interest. We report the occurrence of apoptosis in cultured insect cells and mammalian depleted of nutrients or induced by chemical and viral agents, and supplementation of these cultures with hemolymph of Lonomia obliqua is able to extend the viability of the culture avoiding death by apoptosis. This study aimed to determine in advance the anti-apoptotic mechanism of action of a protein isolated from hemolymph of L. obliqua. The identification of this mechanism may help develop strategies for controlling apoptosis in cell culture allowing the optimization of productivity of viral and recombinant proteins.
 
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Publishing Date
2012-03-01
 
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