光核桃叶片蛋白双向电泳技术的完善  

Improvement of two-dimensional gel electrophoresis for leaf blade proteins of Prunus mira( Amygdalus mira Koehne Kov.et Kost)

作　　者：曹原[1] 周宇航[1] 王忠奎[2] 孟凡娟[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040 [2]西藏大学农牧学院,西藏林芝860000

出　　处：《黑龙江畜牧兽医》2017年第3期129-131,294,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31160386)

摘　　要：为了找到如何提取高质量光核桃叶片蛋白质的方法,试验采用三种方法对叶片总蛋白质进行提取,并采用Bradford法进行蛋白质浓度的测定,最后进行双向凝胶电泳,通过比较蛋白质浓度以及蛋白质斑点数量及分布特点来确定最佳方法。结果表明:对样品研磨的精细程度、丙酮清洗沉淀的次数等操作细节均能影响提取结果;用醋酸铵-甲醇提纯后的电泳结果最佳。说明醋酸铵-甲醇法可应用于光核桃叶片蛋白质组学的研究。To find a way how to extract high - quality leaf blade protein of Prunus mira, the total protein of the leaf blades was extracted using three methods, including TCA- acetone method, optimized TCA -acetone method, and ammounium acetatemethanol method. The Bradford method was used for the determination of protein concentrates, and then two - dimensional gel electrophoresis was conducted in the test. The optimal method was determined through comparing the protein concentration, the number and distribution characteristics of protein spots. The results showed that some operational details could affect the extraction results, including the grinding degree of samples, the number of precipi- tation washed with acetone, and so on. The electrophoresis result was optimal after using ammounium acetate - methanol for the purification. The result indicates that the ammounium acetate - methanol method can be applied to the study of proteomics in the leaf blades of Prunus mira.

关 键 词：光核桃 蛋白质提取 TCA-丙酮法 醋酸铵甲醇法 双向电泳 

分 类 号：S664[农业科学—果树学] O629.73[农业科学—园艺学]