Submitted to: Journal of Chromatography A
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/3/2005
Publication Date: 5/3/2005
Citation: Zukas, A.A., Breksa III, A.P. 2005. Extraction Methods for Analysis of Citrus Leaf Proteins by Two-Dimensional Electrophoresis. Journal of Chromatography A. V. 1078:201-205. Interpretive Summary: Human encroachment on land traditionally used for citrus cultivation, environmental changes, emerging diseases and pests, in addition to changing consumer preferences have driven both researchers and commercial producers of citrus to search for methodologies to reduce the time and cinancial costs in generating new varieties. A proteomics approach using two-dimensional electrophoresis analysis in combination with mass spectrometry has the potential to be a powerful tool in the selection and evaluation of new varieties. However, realization of the full potential of two-dimensional electrophoresis (2-DE) is dependent on good sample preparation. We have developed general procedures for the extraction of Citrus leaf proteins for analysis by 2-DE.
Technical Abstract: General procedures for the extraction of citrus leaf proteins for analysis by two-dimensional electrophoresis (2-DE) were developed through the evaluation and modification of existing methods. Among the methods evaluated, the best results were obtained when Tris-NCI, KCI, and phenol extractions were followed by precipitation with organic solvents to purify and concentrate the samples. The utility of the Tris-NCI extraction method was demonstrated on the leaves of six genetically different Citrus varieties. The methods described are versatile and result in high resolution 2-DE gels of Citrus leaf proteins.