Expressão e purificação de proteínas recombinantes do organismo modelo Neurospora crassa

• Autor(a) principal: Matsuno, Guilherme Eiji [UNESP]
• Data de Publicação: 2014
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/142915; http://www.athena.biblioteca.unesp.br/exlibris/bd/capelo/2016-07-08/000867533.pdf
• topic: Neurospora crassa; Proteínas recombinantes; Dicroismo circular; Analise cromatografica; Proteinas - Pesquisa; Circular dichroism

Model organisms are those considered representatives to fundamental attributes of the kingdoms of organisms or even life itself. Among model organisms, the early works of George W. Beadle and Edward L. Tatum with the fungus Neurospora crassa stimulated the use of microorganisms and has initiated a new era, combining genetics and biochemistry. The fungus N. crassa is easy to grow, showing greater morphological and developmental complexity due to its multicellular characteristics. Moreover, many proteins are predicted as similar proteins in animals, plants and other filamentous fungi. This information emphasizes the potential of N. crassa as a model for elucidation of still unknown biochemical and genetic mechanisms. However, 41% of the analyzed proteins are related to known proteins. Another 30% are hypothetical proteins with no similarity to proteins deposited in databases. Among metabolic pathways targets of studies in Neurospora, an interesting feature is the glycogen metabolism. Under stress conditions, the temperature (heat shock), the glycogen concentration decreases, while in yeast the behavior is opposite. Thus, studies have identified proteins involved in these pathways. The Importin-α protein is important because of its role in recognizing proteins to be transported from the cytoplasm to the cell nucleus. Its study is important as it participates in the transport of proteins involved in the regulation of glycogen metabolism. Another protein is NCU03482, which is identified as a helicase RuvB-like protein, which is responsible for the remodeling of chromatin in human protein. In this work, the expression and purification of recombinant protein Importin-α (Imp-α) and NCU03482 were performed, and the circular dichroism experiment with the sample of Imp-α. Results indicate that the proteins were expressed in Escherichia coli successfully, as well as purification of them. However, among the proteins, the Imp-α was obtained in a...