Evaluation of protein extraction methods for Vitis vinifera leaf and root proteome analysis by two-dimensional electrophoresis.
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
Neila
N
Centre de Biotechnologie de Borj Cédria, Tunisia. jel_neil@yahoo.fr
Salem
Asma Ben
AB
Ghorbel
Abdelwahed
A
Jouira
Hatem Ben
HB
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000302:isolation & purification*; D018517:Plant Roots / Q000737:chemistry*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D027843:Vitis / Q000737:chemistry*
D010940:Plant Proteins; D020543:Proteome