芒果叶片双向电泳体系的建立及优化  

Establishment and Optimization of 2-DE Technique System in Leaf Proteome of Mango

作　　者：朱飞冀 詹儒林[2] 柳凤[2] 李国平[2] 赵艳龙[2] 常金梅[2] 何衍彪[2] 

机构地区：[1]海南大学环境与植物保护学院,海口570228 [2]中国热带农业科学院南亚热带作物研究所,湛江524091

出　　处：《基因组学与应用生物学》2017年第2期814-819,共6页Genomics and Applied Biology

基　　金：中央级非盈利性研究机构基本科研业务费研究专项(1630062014007);海南自然科学基金(20163109)共同资助

摘　　要：为建立适用于芒果叶片总蛋白的双向电泳(two-dimensional electrophoresis,2-DE)体系,使用3种方法(尿素/硫脲法,酚法和三氯乙酸/丙酮法)提取芒果叶片蛋白,采用2种水化方式(主动水化和被动水化)和4种样品上样量(1 000μg,1 300μg,1 500μg,1 800μg)进行双向电泳,G-250考染后,通过PDQuest软件对双向电泳图谱进行比较分析。结果表明,三氯乙酸/丙酮法提取效果最好,所得单向SDS-PAGE条带数与2-DE蛋白点数均多于其它2种方法;主动水化对高丰度蛋白点聚焦效果较好;1 500μg蛋白上样量电泳图谱分辨率最高。该方法的建立为开展芒果蛋白质组学研究提供了参考。In order to establish a proper two-dimension gel electrophoresis （2-DE） protocol for proteomic study of mange leaves, three extraction methods （ureaJthiourea extraction, phenol extraction and TCA/acetone precipitation methods） were appliedtothe extraction of mango leaf protein, two types of rehydration ways （active rehydration and passive rehydration）and three kinds of loading quantityofsample （1 000 μg, 1 300 μg, 1 500 μg, 1 800 μg）were designed. After 2-DE electrophoresis, G-250 dyeing, the 2-DE maps were analyzed with PDQuest software. The results showed that TCA/acetone was the most suitable extraction method, the number of protein bands in the SDS-PAGE and protein spots in the 2-DE maps were more than the other two methods; The better high abundance protein spots isoelectric focusing result was obtained by active hydration; The optimum resolution and the quality of the 2-DE maps were improved for proteins loading amount of 1 500μg. The establishment of this method might be practical for the mango proteomics.

关 键 词：芒果 叶片 蛋白提取 双向电泳 

分 类 号：S667.7[农业科学—果树学]