Applied and Establishment of Two-Dimensional Polyacrylamide Gel Electrophoresis System for Anther Proteome of Lycium barbarum

Using the method of modified TCA-acetone combined with Tris-HCl,Lycium barbarum anther proteins were extracted.Composition of protein lysate,the pH range of IPG strip,sample loading quantity and staining methods were explored.The results showed that reproducible and high-quality 2-DE maps of anther protein can be obtained with 17 cm IPG strip,400 μg of sample loading quantity per strip,lysis buffer containing 2 mol/L thiourea and nitrate silver staining,and anther protein spots mainly distributed within the pH 4~7 range.Using this system,we studied metaphase anther proteins of 'Ningqi 1'and 'Ningqi 2'.About 500 proteins were detected on 2-DE gels of pH 4~7 by PDQuest 8.0 software.Among them,25 differently at least 2-fold expressed spots were found.