Protein extraction from Phoenix dactylifera L. leaves, a recalcitrant material, for two-dimensional electrophoresis.
This work was aimed at optimizing a protein extraction procedure for date palm (Phoenix dactylifera L.) leaves, a highly recalcitrant plant tissue for 2-DE. Five protein extraction protocols based on different protein precipitation agents (TCA/acetone vs. phenol (Ph) methods) and protein resolubilization methods (physical treatments, e.g., sonication, shaking and/or heating) were tested. Ph/SDS extraction with methanol/ammonium acetate precipitation, followed by DOC preincubation and TCA/acetone precipitation and, finally, solubilization by shaking in rehydration solution was found to be the best protein extraction method. We conclude that DOC with TCA/acetone precipitation step eliminates interfering compounds, thus allowing efficient resolubilization of date palm leaf proteins. This method could be appropriate for proteomic studies such as date palm colonization by entomopathogenic fungi.
Department of Marine Sciences and Applied Biology, University of Alicante, Alicante, Spain. sonia.gomez@ua.es
Sonia
S
Gómez-Vidal
Manuel
M
Tena
Luis Vicente
LV
Lopez-Llorca
Jesús
J
Salinas
D028023:Arecaceae / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results
D010940:Plant Proteins