果梅花芽蛋白质双向电泳优化体系的建立  

Optimization of two-dimensional electrophoresis conditions for floral buds proteome analysis of Japanese apricot

作　　者：庄维兵[1] 高志红[1] 章镇[1] 侍婷[1] 邵静[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095

出　　处：《南京农业大学学报》2011年第6期47-52,共6页Journal of Nanjing Agricultural University

基　　金：农业部公益性行业(农业)科研专项经费(201003058);江苏省自然科学基金项目(BK2011642)

摘　　要：以果梅花芽为材料,比较了TCA/丙酮法和酚抽法2种总蛋白质提取方法对蛋白质双向电泳(2-DE)的影响,同时对IPG胶条种类、凝胶浓度、蛋白质上样量及染色方法等环节进行了优化。结果表明:TCA/丙酮法比酚抽法在2-DE凝胶图谱上的蛋白质点数增多,图谱背景也比较清晰,没有明显的纵横条纹;采用17 cm pH 4～7的胶条、10%的凝胶浓度、1.5～1.8 mg的蛋白质上样量,经改良胶条考马斯亮蓝G-250染色后可获得背景清晰、重复性好、蛋白质分辨率高的2-DE图谱。高质量的果梅花芽蛋白质提取方法和蛋白质双向电泳体系的优化可为果梅蛋白质组学研究奠定技术基础。The effects of different protein extraction methods on 2-DE maps were compared and the loading quantity of sample,the gel concentration and the staining methods were also optimized for the analysis of 2-DE system in Japanese apricot.The results showed that the samples prepared by TCA/acetone precipitation had been observed with more protein spots in the 2-DE maps than those prepared by phenol extraction method,clearer background and slighter streaks.And a high quality 2-DE map with low background was obtained using the following optimized procedure：loading 1.5-1.8 mg protein sample on 17 cm IPG strip with pH 4-7,SDS-PAGE with 10% gel concentration,and finally detecting proteins with modified colloidal coomassie brilliant blue G-250 staining.The optimized system would facilitate the proteomics research in Japanese apricot.

关 键 词：果梅 花芽 蛋白提取 双向电泳 

分 类 号：S662.4[农业科学—果树学]