African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12191

Full Length Research Paper

Two-dimensional profiling of Xanthomonas campestris pv. viticola proteins extracted by four different methods

Myrzânia de Lira Guerra*
  • Myrzânia de Lira Guerra*
  • Departamento de Agronomia/Fitossanidade, Universidade Federal Rural de Pernambuco - UFRPE, 52171-270, Recife, PE, Brazil.
  • Google Scholar
Carolina Barbosa Malafaia
  • Carolina Barbosa Malafaia
  • Departamento de Bioquímica, Biologia Molecular, Universidade Federal de Pernambuco, Av. Professor Moraes Rego, 1235, Cidade Universitária, Recife, 50740-560, Brazil.
  • Google Scholar
Túlio Diego da Silva
  • Túlio Diego da Silva
  • Departamento de Bioquímica, Biologia Molecular, Universidade Federal de Pernambuco, Av. Professor Moraes Rego, 1235, Cidade Universitária, Recife, 50740-560, Brazil.
  • Google Scholar
Rosa de Lima Ramos Mariano
  • Rosa de Lima Ramos Mariano
  • Departamento de Agronomia/Fitossanidade, Universidade Federal Rural de Pernambuco - UFRPE, 52171-270, Recife, PE, Brazil.
  • Google Scholar
Márcia Vanusa da Silva
  • Márcia Vanusa da Silva
  • Departamento de Bioquímica, Biologia Molecular, Universidade Federal de Pernambuco, Av. Professor Moraes Rego, 1235, Cidade Universitária, Recife, 50740-560, Brazil.
  • Google Scholar
Elineide Barbosa de Souza
  • Elineide Barbosa de Souza
  • Departamento de Biologia/Microbiologia - UFRPE, 52171-030, Recife, PE, Brazil.
  • Google Scholar


  •  Received: 30 May 2014
  •  Accepted: 04 August 2014
  •  Published: 20 August 2014

Abstract

An efficient method for protein extraction is a prerequisite for the successful implementation of proteomics, which is being used as a tool in the study of the interaction between plants and phytopathogens. With the objective to optimize a method of protein extraction for proteomic analysis of the phytobacterium Xanthomonas campestris pv. viticola, the efficiency of four methodologies were compared, based on the two-dimensional gel electrophoresis profile (2D-PAGE). Trizol®, phenol, centrifugation and lysis methods were tested and through quantitative and qualitative analysis, the most suitable method to obtain high-quality protein was selected. All methodologies enabled the extraction of a significant amount of proteins; nevertheless, the centrifugation method allowed obtaining the highest concentration of solubilized proteins. However, the analysis of the 2D-PAGE gel images revealed a larger number of spots in the lysis method when compared to the others. Taking into consideration the quality of the results and the practical advantages of the lysis method, this is recommended as the best option for total protein extraction of X. campestris pv. viticola for proteomic studies.

 

Key words: Bacterial canker, Vitis vinifera, proteomics, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2D-PAGE).