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Di Ciero,Luciana; Bellato,Cláudia de M.; Meinhardt,Lyndel W.; Ferrari,Fernanda; Castellari,Rafael R.; Marangoni,Sérgio; Novello,José C.. |
Four different detergents, ASB 14, SB 3-10, CHAPS and Triton X100, were utilized to determine the optimal detergent for the solubilization of membrane proteins from the phytopathogenic bacterium Xylella fastidiosa. These proteins were differentially solubilized in distinct buffers containing the detergent and subjected to bidimensional electrophoresis within the non-linear pH range of 3-10. The detergents ASB 14 and SB 3-10 were the most effective revealing 221 and 157 spots, respectively. CHAPS and Triton X100 were less effective and revealed only 72 and 43 spots, respectively. MALDI-TOF tryptic peptide mass fingerprinting of 18 excised proteins from the ASB 14 treatment revealed that 83% were membrane proteins and that the theoretical efficiency of... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: X. fastidiosa; 2-DE; Extraction; Proteome; Solubilization. |
Ano: 2004 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200018 |
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Bellato,Cláudia de M.; Garcia,Andréia K.M.; Mestrinelli,Fabiana; Tsai,Siu M.; Machado,Marcos A.; Meinhardt,Lyndel W.. |
An in vitro system was developed to induce and identify Xylella fastidiosa proteins that were differentially expressed in the presence of callus-derived extracts from its host, the citrus cultivar Pêra. To optimize the induction, we first developed a single culture medium for the growth of both, host and bacteria. This medium, CPXPm7, which mimics the citrus xylem sap, showed that X. fastidiosa at 72 h post-incubation had 10(8) colony forming units mL-1, while Pêra cells had the highest fresh weight content (0.79 g). After testing various methods of co-cultivation of the bacteria and host callus grown in this single medium, the best induction procedure was to grow X. fastidiosa in a solid medium amended with an extract of Pêra callus grown in CPXPm7.... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: 2DE; Callus; Pathogenicity; Protein; Xylella-citrus interaction. |
Ano: 2004 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822004000200012 |
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