油菜种子微粒体制备及蛋白浓度检测方法研究

目的]建立利用差速离心法制备油菜种子微粒体的方法及微粒体蛋白浓度测定的紫外分光光度检测法。方法]分别取授粉后47 d的高油酸与低油酸材料新鲜角果,将种子剥出,应用差速离心法提取油菜种子微粒体,采用Bradford法测定种子微粒体蛋白浓度。结果]离心管底可见沉淀,牛血清白蛋白标准曲线范围为0～500μg/ml,回归方程为：Y=0.001 34X＋0.064（r=0.996 3）,日内和日间精密度分别为3.52%、1.55%、0.54%以及7.51%、1.99%、0.68%。油菜种子微粒体总蛋白质含量为高油酸材料406μg/g,低油酸材料335μg/g。结论]利用差速离心法制备得到高质量油菜种子微粒体蛋白,可为进一步研究油菜种子油酸脱氢酶活性提供依据。

Study on Minute Particles Preparation and Methodology of Protein Concentration in Rapeseeds

Objective]The aim was to establish the method of rapeseeds minute particles preparation by using different velocity centrifugation and the UV determination methodology of protein concentration in minute particles of rapeseed.Method] Both high oil-content materials and low oil-content materials of developing silique were collected at 47 days after pollination.The silique was peeled,and minute particles of rapeseeds were extracted by different velocity centrifugation.The Bardford method was used to measure ...