Evaluation of Protein Extraction Methods for Vitis vinifera Leaf and Root Proteome Analysis by Two-Dimensional Electrophoresis
Neila Jellouli , Asma Ben Salem, Abdelwahed Ghorbel and Hatem Ben Jouira
Laboratoire de Physiologie Mol¨¦culaire de la Vigne, Centre de Biotechnologie de Borj C¨¦dria, BP 901- Hammam Lif 2050, Tunisia
Correndence for author Tel.: +216 98 93 79 81; E-mail: jel_neil@yahoo.fr
Online on 16 June 2010 at www.jipb.net and www.interscience.wiley.com/journal/jipb 10.1111/j.1744-7909.2010.00973.x
Abstract
An efficient protein extraction method is crucial to ensure successful separation by two-dimensional electrophoresis (2-DE) for recalcitrant plant species, in particular for grapevine (Vitis vinifera L.). Trichloroacetic acid-acetone (TCA-acetone) and phenol extraction methods were evaluated for proteome analysis of leaves and roots from the Tunisian cultivar 'Razegui'. The phenol-based protocol proved to give a higher protein yield, a greater spot resolution, and a minimal streaking on 2-DE gels for both leaf and root tissues compared with the TCA-based protocol. Furthermore, the highest numbers of detected proteins on 2-DE gels were observed using the phenol extraction from leaves and roots as compared with TCA-acetone extraction.
Jellouli N, Ben Salem A, Ghorbel A, Ben Jouira H (2010) Evaluation of protein extraction methods for Vitis vinifera leaf and root proteome analysis by two-dimensional electrophoresis. J. Integr. Plant Biol. doi: 10.1111/j.1744-7909.2010.00973.x
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Received 8 Dec 2009 Accepted 27 May 2010
© 2009 Institute of Botany, Chinese Academy of Sciences
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