豌豆白化苗蛋白质双向电泳技术的建立  

Establishment of Two-dimension Electrophoresis Technique in Pea (Pisum sativum) Albino Seedlings

作　　者：赵鑫[1,2] 刘红霞[2] 陈佰鸿[1] 毕阳[1] 

机构地区：[1]中国农业科学院作物科学研究所农业部作物遗传育种重点开放实验室国家农作物基因资源与基因改良国家重大科学工程,北京100081 [2]甘肃农业大学农学院,兰州730070

出　　处：《生物技术通报》2010年第10期100-103,共4页Biotechnology Bulletin

基　　金：国家自然科学基金(30500342);中国博士后基金(2004036176)

摘　　要：以豌豆白化苗为试验材料,分析了不同的蛋白质提取时间(4 h、8 h及过夜)和蛋白质上样量(500、1 000、2 000μg)对双向电泳蛋白质得率和等电聚焦效果的影响。结果表明,不同的蛋白提取时间和蛋白上样量对等电聚焦效果影响较大。适宜的豌豆白化苗提取时间应至少长于8 h,最好提取过夜,能裂解得到大量蛋白质;适宜的豌豆白化苗蛋白上样量为1 000μg,此条件下,双向电泳的分辨率最高,分离出的蛋白大多为小分子量功能性蛋白。This study focused on the specific problems of protein extraction from pea seedlings tissues. Sample preparation is a critical step in a two-dimensional gel electrophoresis proteome approach and is absolutely essential for good results. We used different extract time （4 h,8 h,and overnight） and protein loading amount （500,1 000,2 000 μg） to analyze the effects of these two factors on the results of 2-DE. Results showed that the optimal protein extraction time for pea albino seedlings is not lesser than 8 hours, preferably overnight,which can crack a large number of proteins;optimal protein loading amount is 1 000μg,which can obtain as many functional proteins as possible and the best resolution in two-dimensional eleetrophoresis. The establishment of 2-DE in pea Mbino seedlings laid a basis for further study the function of protein involved in pea defense response.

关 键 词：蛋白质组学 豌豆 2-DE 

分 类 号：Q946[生物学—植物学]