title: Selection of a protein solubilization method suitable for phytopathogenic bacteria: a proteomics approach.
abstract: BACKGROUND
  Finding the best extraction method of proteins from lysed cells is the key step for detection and identification in all proteomics applications. These are important to complement the knowledge about the mechanisms of interaction between plants and phytopathogens causing major economic losses. To develop an optimized extraction protocol, strains of Acidovorax citrulli, Pectobacterium carotovorum subsp. carotovorum and Ralstonia solanacearum were used as representative cells in the study of phytopathogenic bacteria. This study aims to compare four different protein extraction methods, including: Trizol, Phenol, Centrifugation and Lysis in order to determine which are more suitable for proteomic studies using as parameters the quantity and quality of extracted proteins observed in two-dimensional gels.
  
  
  RESULTS
  The bacteria studied showed different results among the tested methods. The Lysis method was more efficient for P. carotovorum subsp. carotovorum and R. solanacearum phytobacteria, as well as simple and fast, while for A. citrulli, the Centrifugation method was the best. This evaluation is based on results obtained in polyacrylamide gels that presented a greater abundance of spots and clearer and more consistent strips as detected by two-dimensional gels.
  
  
  CONCLUSIONS
  These results attest to the adequacy of these proteins extraction methods for proteomic studies.
authors: Programa de Pós-Graduação em Ciências Biológicas - Centro de Ciências Biológicas - Universidade Federal de Pernambuco, Rua Prof. Nelson Chaves s/n, Cidade Universitária, CEP 50670-901 Recife, PE Brasil.
authors: Carolina B
authors: CB
authors: Malafaia
authors: Programa de Pós-Graduação em Fitopatologia - Universidade Federal Rural de Pernambuco, Av. Dom Manoel de Medeiros, s/n - Dois Irmãos, CEP: 52171-900 Recife, PE Brasil.
authors: Myrzânia L
authors: ML
authors: Guerra
authors: Programa de Pós-Graduação em Ciências Biológicas - Centro de Ciências Biológicas - Universidade Federal de Pernambuco, Rua Prof. Nelson Chaves s/n, Cidade Universitária, CEP 50670-901 Recife, PE Brasil.
authors: Túlio D
authors: TD
authors: Silva
authors: Departamento de Bioquímica, Universidade Federal de Pernambuco, Rua Prof. Moraes Rego s/n, Cidade Universitária, 50670-420 Recife, PE Brasil.
authors: Patrícia Mg
authors: PM
authors: Paiva
authors: Departamento de Biologia, Universidade Federal Rural de Pernambuco, Av. Dom Manoel de Medeiros, s/n - Dois Irmãos, CEP: 52171-900 Recife, PE Brasil.
authors: Elineide B
authors: EB
authors: Souza
authors: Departamento de Bioquímica, Universidade Federal de Pernambuco, Rua Prof. Moraes Rego s/n, Cidade Universitária, 50670-420 Recife, PE Brasil.
authors: Maria Ts
authors: MT
authors: Correia
authors: Departamento de Bioquímica, Universidade Federal de Pernambuco, Rua Prof. Moraes Rego s/n, Cidade Universitária, 50670-420 Recife, PE Brasil.
authors: Márcia V
authors: MV
authors: Silva
keywords: Acidovorax citrulli; Pectobacterium carotovorum subsp. carotovorum; Proteome analysis; Ralstonia solanacearum; Two-dimensional gel electrophoresis