ISHS Acta Horticulturae 728: V International Congress on Cactus Pear and Cochineal
A 2D-PAGE ANALYSIS OF PROTEIN DEPOSITION DURING THE DEVELOPMENT OF PHYLLOCLADS OF OPUNTIA FICUS-INDICA |
| Authors: | E.A..R Vasconcelo, P.A.S. Souza, R.M. Liamoca Zarate, F.A.P. Campos |
| Keywords: | cactus pear, forage crops, bi-dimensional electrophoresis, protein deposition, Cactaceae |
| Abstract:
A method for the extraction and analysis of proteins by two-dimensional electrophoresis from phylloclads of the cactus pear was developed. This method is based on the extraction of proteins from freeze-dried phylloclads in 50 mM pyridine, 10 mM thiourea, 1% SDS, pH 5.0, and precipitation of the extracted proteins with 10% TCA in acetone. Proteins were separated in the first dimension by isoelectric focusing in Immobiline DryStrips, pH 3-10. The second dimension of electrophoresis was performed in a uniform 15% separating gel at 25ºC and a under constant current of 25 mA. The efficiency of this protocol was demonstrated by comparison of the cactus pear proteome at four developmental stages of the phylloclads. | |
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