知识与财富的链接
为探索适合小麦穗突变体Sda1叶片蛋白的双向电泳体系,寻找Sda1与其野生型叶片的差异蛋白,以Sda1与其野生型的抽穗期叶片为材料,从蛋白质提取、双向电泳条件等方面对适合Sda1及其野生型叶片蛋白的双向电泳体系进行探索。分析试验得到的双向电泳图谱,发现利用TCA/丙酮提取叶片总蛋白,用pH 4~7的17cm线性胶条,采用13%浓度的分离胶,上样量为900μg,利用改良的等电聚焦程序,能够得到背景清晰、分辨率高的电泳图谱;电泳图谱在低分子区蛋白点分布均匀,并且重复性好。利用PDQuest 8.0.1软件分析图谱,得到27个差异点蛋白,相比于野生型植株,Sda1突变体表现为蛋白表达量下调与缺失,发现6个缺失蛋白,21个表达下调蛋白。
In order to explore suitable 2-DE(two-dimensional electrophoresis) system for Sda1(spike development atrophy 1) leaf protein to screen differential protein expression between Sda1 and its wildlife type in wheat(Triticum).The protein extraction method and two-dimensional electrophoresis conditions were studied. A clear background, well distributed protein spots and reproducible 2-DE maps can be obtained by using the TCA/acetone protein extraction method, with 13% separation gel, a sample amount of 900 μg, the linear strip 17 cm immobilized pH gradient(IPG) strip pH 4 to 7, modified isoelectric focusing(IEF) process, and modified Coomassie Brilliant Blue for staining. Using this optimized 2-DE system, some differential protein spots between Sda1 and its wildlife-type were found. Compared with its wildlife-type,21 spots were down-expressed, and six spots were not expressed in Sda1.
