麦醇溶蛋白A-PAGE方法的优化和改进

麦醇溶蛋白A-PAGE方法的优化和改进 Improvement of the Standard Method of Acid Polyacrylamide Gel Electrophoresis（A-PAGE） Recommended by ISTA for Hordeins and Gliadins 邵锦震 SHAO Jin\|zhen 1 first-author 丁毅 DING Yi 1 武汉大学植物发育生物学教育部重点实验室,湖北,武汉,430072 武汉大学植物发育生物学教育部重点实验室,湖北,武汉,430072 Key Laboratory of MOE for Plant Development Biology, Wuhan University, Wuhan 430072,Hubei, China) \ \ Key Laboratory of MOE for Plant Development Biology, Wuhan University, Wuhan 430072,Hubei, China) \ \ 以大麦、小麦品种等为材料 ,在 ISTA的标准电泳程序基础上分别进行浓度为 10 % ,12 % ,14% ,16 % ,18%和 2 0 %的酸性聚丙烯酰胺 （ A- PAGE）均一胶电泳来分离种子的麦醇溶蛋白 ,并与 10 %～ 2 0 %的线性梯度A- PAGE分离效果进行比较 .经优化和改进 ,结果表明 :18%的 A- PAGE方法效果最佳 ,既具有 ISTA推荐的A- PAGE方法制胶简便、电泳快速、重复性好的优点 ,又具有线性梯度 A- PAGE方法分辨率高的特点 ,尤其是在小分子量的快带区 ,电泳成带明显 ,达到梯度胶的分离效果 .在供试的大麦、小麦品种中 ,电泳图谱总条带数比 ISTA推荐的 10 %均一胶所得电泳图谱条带数分别增加 1～ 8条和 2～ 7条 .该方法既适用于大麦、也适用于小麦醇溶蛋白的分离 .并对 18%的 A- PAGE方法的适用范围等进行了讨论 Hordeins and gliadins were separated with different gel concentration such as 10%、12%、14%、16%、18%、20%, On the basis of the standard method of acid polyacrylamide gel electrophoresis （A\|PAGE） . The comparison was made for separation effects on the alcohol\|soluble proteins between the methods of ISTA and 10% 2 0% linear gradient A\|PAGE. By means of improvement of the standard method of ISTA （10%）, the results revealed that the best resolution could be obtained with 18% gel concentration. With the advantages of reproducibility, time saving and simple operation, the improved method is similar to that recommended by ISTA. On the other hand, high\|resolution can be obtained by this technique （18%）as that in the 10% 2 0% linear gradient A\|PAGE. This method separated more polypeptides, especially in fast\|band region of the electrophoregram. There were 1 8 and 2 7 more bands in the electrophoretic patterns for hordeins and gliadins respectively with 18% A\|PAGE than that with 10% A\|PAGE. And, the appling range of the improved method was discussed.\; 大麦醇溶蛋白 小麦醇溶蛋白 A-PAGE 电泳图谱 分辨率 10.14188/j.1671-8836.2000.06.021 Q946.1 国家自然科学基金资助项目! （ 3 9670 0 5 1）;教育部骨干教师计划项目资助 2000-06-01 2021-04-01 6