玉米叶片总蛋白提取和双向电泳技术的改进  

Two Dimensional Electrophoresis and Extraction of Total Protein of Maize

作　　者：李冠军[1] 付凤玲[1] 

机构地区：[1]四川农业大学玉米研究所西南作物基因资源与遗传改良教育部重点实验室,四川雅安625014

出　　处：《玉米科学》2006年第6期100-103,共4页Journal of Maize Sciences

基　　金：国家自然科学基金项目(30571172);教育部长江学者和创新团队发展计划(IRT0453)

摘　　要：针对植物组织蛋白质含量较低,有些组分在两性电解质中的溶解性低以及绿色组织中的色素、酚类、醌类等次生代谢产物干扰蛋白质双向电泳分离效果的问题,以玉米苗期叶片为材料,对蛋白质裂解液、蛋白质浓度测定体系、等电聚集电泳电压和时间、聚丙烯酰胺凝胶浓度等技术参数做了改进。用改进的裂解液提取的蛋白质样品浓度达3.8μg/μL,比改进前提高90%。通过在浓度测定体系中加入HCl并用裂解液代替超纯水配制标准蛋白,可得到线性更好的蛋白浓度测定标准曲线。用改进后的等电点聚焦电泳的电压、时间和聚丙烯酰胺凝胶浓度,可从玉米叶片总蛋白质样品中分离出963个清晰的蛋白点,在其他条件相同的情况下,比改进前的技术体系多分离出429个蛋白点。This study aimed at the problem of low content of protein in plant tissue, low solubility of some components in amphoteric electrolytes and the disturbance of secondary metabolites such as chromatophore, phenols and quinones to the separation of two-dimensional electrophoresis. Using leaves of maize in seedling as material, index such as biolytics, concentration measurement system, condition of isoelectric focusing and the density of polyacrylamide gel were improved. Though comparison, the concentration of protein extracted with the improved biolytics was as high as 3.8 μg/μL, 90% than before, better linearity was obtained and proteins were better separated on two-dimension gels with 963 spots clearly detected, 429 more than before.

关 键 词：玉米 叶片 蛋白质 双向电泳 

分 类 号：S513.01[农业科学—作物学]