CAJ | 학술논문

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。
위료획득순화、완정적옥미협록체단백질,이옥미품충정단958위시험재료,이용차속리심여자당밀도제도법분리협록체,병대기순도화완정도진행검측；통과광학현미경직접관찰협록체적형태결구병측정과양화경매적활성급Hill반응속솔,증명이분리출순도교고、완정적협록체。수후채용2충불동적단백질제취방법분별제취옥미협록체총단백질,병용Bradford법진행단백질정량, SDS-PAGE전영결과표명,TCA병동침정법대옥미협록체단백질적제취불충분,이TCA병동여분추제상결합법명현비TCA병동침정법효과호,제출적협록체단백질적조대수목다,봉도교고,병득도료교이상적옥미협록체쌍향전영도。
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 de-tected. The maize chloroplast proteins were extracted using two different methods and quantified by Brad-ford method. The SDS-PAGE electrophoresis results showed that the extraction of maize chloroplast pro-teins 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 two-dimensional electrophoresis figure was perfect.