Eletroforese em gel de poliacrilamida (SDS-PAGE) e analise numerica de leveduras isoladas da cavidade oral de humanos

AUTOR(ES)

Edvaldo Antonio Ribeiro Rosa

DATA DE PUBLICAÇÃO

1997

RESUMO

This work aimed to evaluate distincts methodologies for whole-cell proteins extraction of yeasts isolated from oral cavity, for application in polyacrylamide gel electrophoresis (SDS-PAGE). Such proteins were obtained by disruption with glass beads, trichloroacetic acid action, loading sample buffer action, sonication and cellular cryofracture by liquid nitrogen and applied in polyacrylamide gel electrophoresis. The profiles were analised by area integralization densitometry and showed that the best electrophoretic profile for proteins were obtained by cellular cryofracture and sonication. Glass beads and loading sample buffer showed less satisfactories results and the trichloroacetic acid method was not able to disclose applicable proteins bands to be used in studies of relationships of Candida species. The experimental results let to the observation that the species studied by using liquid nitrogen (cellular cryofracture method) showed the best electrophoretic profiles when compared to other metodologies. At the second part of this work, the whole-cell proteins were obtained by cellular cryofracture, separated by SDS-PAGE in a discontinous gel system and analised by densitometry of peaks detection. The migration distance values (Rfs) were converted in binominal data that after computer-process, gave the phenetic similarities among the different strains within the same species and among the different species within the same genera. The results obtained alowed to cluster the different strains and species by their phenetic similarities, giving clusters composed uniquely by specimen of the same species. This result suggests that the numerical analysis of characteristics based on electrophoregrams of whole-cell proteins of oral yeasts is a tool with great discriminately potential in studies of identification, sistematics and biodiversity

ASSUNTO(S)

analise numerica - processamento de dados candida eletroforese

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