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African Journal of Biotechnology

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

M de Lira Guerra, CB Malafaia, TD da Silva, R de Lima Ramos Mariano, MV da Silva, EB de Souza

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.

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

African Journal of Biotechnology, Vol 13(31) 3531-3537



http://dx.doi.org/10.5897/AJB2014.13953
AJOL African Journals Online