多种小麦种子储藏蛋白的电泳分析

ANALYSIS OF VARIOUS KINDS OF WHEAT STORAGE PROTEINS WITH ELECTROPHORESIS

采用不同的提取液，对１０个小麦品种的非酶功能性种子储藏蛋白进行提取，分别进行梯度凝胶电泳分析。电泳依据提取液的不同，分别采用酸性或碱性系统。对酸性凝胶催化系统，采用Ａｐ－Ｖｃ－ＦｅＳＯ４系统代替Ｈ２Ｏ２－Ｖｃ－ＦｅＳＯ４系统，克服了酸性凝胶的不足，提高了凝胶的性能并使之容易操作。应用新的催化系统配制的酸性梯度胶，提高了分辨率。并初步尝试以酸性系统分析种子谷蛋白，获得了成功。经过对不同提取液蛋白质的分析，发现除盐溶蛋白品种间存在较小的差异外，其它提取液提取蛋白间存在较大的差异。

Wheat storage proteins were extracted with different solutions.The protein solving in salt solution was resolved in native polyacrylamide gel electrophoresis of Laemmli system,and other proteins were resolved in acid polyacrylamide gel electrophoresis (A PAGE).For A PAGE,a new catalyst system for polyacrylamide gel at low pH has been developed.The system consists of ascorbic acid,ferrous sulfate and ammonium persulfate.In contrast to the hydrogen peroxide catalyst system (ascorbic acid,ferrous sulfate and hydrogen peroxide) widely used under acid condition,the concentrations of ferrous sulfate and ascorbic acid were reduced and ammonium persulfate replaced hydrogen peroxide.The changing improved the gel mechanical property and made the operation easy. Acidic gradient gels were made with new catalyst system and more bands were found.Glutenin subunits were resolved in A PAGE buffered by acetic acid,which contained 5 mol/L urea to maintain protein solubility during migration.Through analysis with electrophoresis,fewer differences in protein solving in salt solution and more in other proteins were found among wheat varieties.