【摘要】 为了获得纯化、完整的玉米叶绿体蛋白质,以玉米品种郑单958为试验材料,利用差速离心与蔗糖密度梯度法分离叶绿体,并对其纯度和完整度进行检测;通过光学显微镜直接观察叶绿体的形态结构并测定过氧化氢酶的活性及Hill反应速率,证明已分离出纯度较高、完整的叶绿体。随后采用2种不同的蛋白质提取方法分别提取玉米叶绿体总蛋白质,并用Bradford法进行蛋白质定量,SDS-PAGE电泳结果表明,TCA丙酮沉淀法对玉米叶绿体蛋白质的提取不充分,而TCA丙酮与酚抽提相结合法明显比TCA丙酮沉淀法效果好,提出的叶绿体蛋白质的条带数目多,丰度较高,并得到了较理想的玉米叶绿体双向电泳图。更多还原

【Abstract】 In order to obtain the pure and intact chloroplast protein,the maize variety Zhengdan 958 was used as experimental material to isolate the chloroplasts by differential centrifugations,and then the intact chloroplasts were purified by sucrose density gradient method. The morphological structure and purity of chloroplasts were observed by optical microscope,and the catalase activity and Hill reaction rate were detected. The maize chloroplast proteins were extracted using two different methods and quantified by Bradford method. The SDS-PAGE electrophoresis results showed that the extraction of maize chloroplast proteins by TCA acetone precipitation method was insufficient. Compared with TCA acetone precipitation,the proteins extracted by TCA acetone and phenol extraction combination were high abundance and their twodimensional electrophoresis figure was perfect.更多还原