油松雌性不育系球果蛋白质双向电泳技术的建立  

Improvement of Two-dimensional Gel Electrophoresis of Proteins from Female Cones of the Female-sterile Pinus tabulaeformis Carr.

作　　者：丁坤善[1] 郑彩霞[1] 包仁艳[1] 姜春宁[1] 

机构地区：[1]北京林业大学生物科学与技术学院,北京100083

出　　处：《植物学通报》2005年第2期190-197,共8页Chinese Bulletin of Botany

基　　金：国家自然科学基金项目"油松雌配子体败育的分子生物学机制"(39970602)

摘　　要：本文建立了油松雌性不育系雌球果蛋白质组研究中的双向电泳技术。第一向采用固定pH值梯度(IPG)胶条在IPGphorTM等电聚焦仪上进行等电聚焦,第二向在恒功率且恒温条件下于Ettan-DALTTMⅡ高通量电泳仪上进行SDS-PAGE电泳,以银染和考马斯亮蓝两种方法染色。通过对全蛋白的提取、胶条pH值和胶条肿胀等技术环节的优化和比较,得到了重复性很高,分离效果良好的蛋白质双向图谱。This paper reported a improvement in 2-D gel electrophoresis of the proteome in female cones of the female-sterile Pinus tabulaeformis Carr. An IPGphorTM unit with immobile pH gradient strips was used in and the EttanTM DALT Ⅱ system under constant power and tempera- ture in SDS-PAGE. The 2-D gel was stained by both methods of coomassie blue and silver. Extraction of proteins, pH of the strips, rehydration of the strips and other parameters were optimized and compared. Higher repeatability and better separating protein pattern could be gained by this technique.

关 键 词：双向电泳技术 不育系 雌性 油松 蛋白质组研究 PAGE电泳 等电聚焦 考马斯亮蓝 恒温条件 技术环节 分离效果 pH值 胶条 雌球果 SDS 电泳仪 高通量 恒功率 重复性 银染 

分 类 号：S791.254[农业科学—林木遗传育种]