新疆沙冬青叶片蛋白质的双向电泳技术研究

Two-dimensional Electrophoresis of Proteins Extraction from Ammopiptanthus nanus(M.Pop.) Cheng f. Leaves

提取新疆沙冬青叶片中蛋白质受到脂类、酚类等次生代谢产物的干扰,针对这种现象,进行了组分提取优化方法和双向电泳技术的改进。实验分析表明,用乙醚淋洗完整叶片,50 mmol.L-1Tris-HCl缓冲液,上样量240μg(7 cm胶条),考马斯亮蓝染色等方法可很大程度增加提取可溶性蛋白含量,结合丙酮-三氯乙酸的沉降,可较完整地提取蛋白质组。

Proteins extraction from leaves of Ammopiptanthus nanus（M.Pop.） Cheng f.is disturbed by some secondary metabolites,such as lipoidal and phenolic substances.To overcome this problem,method of proteins extraction and two-dimensional electrophoresis protocol of Ammopiptanthus.nanus（M.Pop.）Cheng f.were optimized and ameliorated in this paper.Further,study of preparing method,quantitative loading of samples and staining means were conducted.The results show that washing whole leafs with aether,extracting soluble proteins with 50 mmol·L^-1 Tris-HCl（pH 8.5）,combined with acetone-trichloroacetic acid depositing is a good way of extracting proteins from Ammopiptanthus.nanus（M.Pop.）Cheng f.