Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Protein design wikipedia , lookup
Homology modeling wikipedia , lookup
Protein folding wikipedia , lookup
Bimolecular fluorescence complementation wikipedia , lookup
Protein structure prediction wikipedia , lookup
Western blot wikipedia , lookup
Protein purification wikipedia , lookup
Nuclear magnetic resonance spectroscopy of proteins wikipedia , lookup
Identification of soil bacteria belonging to the phylum Acidobacteria by MALDI-TOF mass spectrometry Soares, C. E; Castro, V; Barreto, C; Magalhães, B. S. Programa de Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, DF, Brasil. Recently, it has been shown that bacterial species of the phylum Acidobacteria are abundant in Cerrado soil, representing more than 50% of total 16S rRNA sequences obtained. These bacteria are expected to play an important role in nutrient cycling in this Biome but are very difficult to obtain in culture. Using novel culturing techniques, five Acidobacteria were isolated from soil samples obtained near Cristalina, Goiás. These isolates were identified using 16S rRNA gene phylogeny and characterized by colony and cell morphology. In addition, MALDI-TOF MS has been applied to generate protein profiles of these isolates that enabled the rapid differentiation of new Acidobacteria species. Two protein extraction protocols were tested to optimize the amount of protein obtained. Twenty mass spectra of each protein extract were acquired to ensure the reproducibility of the MS profiles generated the mass range of spectra acquisition was 2000 to 20000 m/z with a Microflex instrument (Bruker Daltonics). Protein profiles were obtained and evaluated by principal component analysis (PCA). These results were in good agreement with classical phylogenetic analysis based upon rRNA 16S sequencing it was possible to validate the application of this methodology for fast identification of species of the Acidobacteria phylum and ensure the reproducibility of the mass spectra obtained used to produce the phylogenetic tree. The use of MALDI-TOF MS for the identification of soil bacteria is dependent upon the generation of a database of protein profiles and these results constitute the first effort towards a unified database for environmental soil bacteria. Keywords: MALDI-TOF MS; Acidobacteria phylum; soil bacteria; Cerrado Support: CNPq, Embrapa . ��������������������������������������������������������������������������� ��������������������������������������������������������������������������������� �����������������������������������������������������