麻竹叶片蛋白质双向电泳体系的建立  

The Establishment of Two-dimensional Gel Electrophoresis System for Leaf Protein of Dendrocalamus latiflorus

作　　者：杨惠琴[1,2] 乔桂荣[2] 蒋晶[2] 刘明英[2] 姜彦成[1] 卓仁英[2] 

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046 [2]中国林业科学研究院亚热带林业研究所,浙江富阳311400

出　　处：《竹子研究汇刊》2012年第3期16-22,27,共8页Journal of Bamboo Research

基　　金：浙江省科技厅重大专项(2010C12010);浙江省自然科学基金项目(R30972340)

摘　　要：以麻竹花药培养再生植株叶片为材料,比较了改良酚抽法、三氯乙酸(TCA)-丙酮沉淀法和裂解液法3种不同方法对麻竹蛋白质双向电泳的影响,并对上样量、等电聚焦程序、平衡时间等进行了探索与优化。结果表明,用改良酚抽法分离麻竹叶片蛋白质、确定上样量为1 mg.IPG胶条-1、等电聚焦总聚焦功率为100 000 W、胶条两步平衡时间均设置为15 min、考马斯亮进行凝胶染色的实验体系可以获得分辨率高、重复性好的双向电泳图谱,凝胶的蛋白斑点匹配率平均为85.57%,总蛋白斑点数的相对标准差为4.86%。本研究建立了一套适合麻竹叶片蛋白质组学分析的双向电泳实验体系,为进一步研究麻竹生长发育分子调控机制及其遗传改良奠定基础。Using the leaves of regenerating bamboos cultured from anther as experimental materials,three kinds of two-dimensional electrophoresis methods,i.e.modified phenol extraction,three acetic acid（TCA）-acetone precipitation and lysis-buffer method,were compared to explore their effectiveness on bamboo proteins.The optimal sample volume,isoelectric focusing procedure and balance time etc.were studied also.The results showed that two-dimensional electrophoresis of high resolution and good repeatability can be obtained when the improved phenol extraction method was used to separate bamboo leaf protein under the following systematic conditions：sample volume of 1 mg·IPG strip-1,isoelectric focusing power of 100 000 W,strip equilibrium time of 15 minutes per step and Coomassie gel dyeing.The gel protein spot matching rate was 85.57% and the relative standard deviation of the total number of protein spots was 4.86%.This study has established a two-dimensional gel electrophoresis experimental system being suitable to the analysis of bamboo leaf protein,and provided a basis for further study on the molecular mechanism of bamboo growth and its genetic improvement.

关 键 词：麻竹 蛋白质 双向电泳 

分 类 号：S795.5[农业科学—林木遗传育种]