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甘蔗叶绿体分离及其蛋白质提取方法研究
作者: 孙 富1  2  黄巧玲1  2  黄 杏1  2  张保青1  2  陈明辉1  2  杨丽涛1  2 *  李杨瑞2  3  4  5 *  
单位: (1 广西大学农学院  南宁 530005;2 亚热带农业生物资源保护与利用国家重点实验室  南宁 530005;3 广西农业科学院  
关键词: 甘蔗  叶绿体  蛋白质  密度梯度离心  
分类号:566.101
出版年,卷(期):页码:2011 ,42 ( 5 ): 页码:463-467
摘要:

【Objective】In order to study sugarcane chloroplasts proteomics, the methods for isolation and purification of intact chloroplast and its protein extraction from leaves of sugarcane seedling were compared. 【Method】The leaves of young seedling of sugarcane variety ROC22 were sampled to isolate the chloroplasts by differential centrifugation. The obtained intact chloroplasts were isolated and purified using sucrose density gradient centrifugation and Percoll gradient centrifugation. The proteins in chloroplast were extracted and analyzed by SDS-PAGE electrophoretic. 【Result】The results indicated that both density gradient centrifugation methods could obtain high rate and high purity of intact chloroplasts, but sucrose density gradient centrifugation saved the time and cost, and resulted in high purity and integrity of chloroplasts. The SDS-PAGE profiles of proteins, separated from the intact chloroplasts, were rich and clear, with high quantity and quality of proteins.【Conclusion】Compared with Percoll gradient centrifugation method, sucrose density gradient centrifugation method was suitable for proteomics studies of sugarcane at subcellular level.

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