Extraction of Total Proteins from Leaves and Peels of Citrus reticulata and Establishment of Two-dimensional Electrophoresis System

Abstract:

The total proteins in peels and leaves of Citrus reticuiata Blanco ‘Chuntianju’ were extracted by four methods, such as Tris-HCl, Thi/Urea, trichloroacetic acid/pacetone precipitation (TCA) and phenol (Phe) extraction methods, respectively, and their effects were compared by protein production, one-way SDS-PAGE and two-dimensional electrophoresis (2-DE). The results showed that TCA method was the best for both leaves and peels of C. reticuiata ‘Chuntianju’. The 2-DE map of proteins extracted by TCA method had high resolution, the protein spots were clear and uniform without stripes, and the number of protein spots were large. It suggested that the TCA method could not only remove the interfering substances in peels and leaves of C. reticuiata ‘Chuntianju’, but also get more stable protein spots.

Key words:Citrus reticuiata;Proteome;Exteaction;Two-dimensional electrophoresis (2-DE)